Fibroblast Cell Counts Research Articles

Dissolving microneedle patches for delivery of amniotic mesenchymal stem cell metabolite products for skin regeneration in UV-aging induced mice

Microneedles offer a promising solution to enhancing dermal delivery of amniotic mesenchymal stem cell metabolite product (AMSC-MP), which contains hydrophilic protein components with high molecular weight, for the purposes of skin rejuvenation and improving human health. This study aimed to evaluate the physicochemical characteristics and in vivo efficacy of AMSC-MP-loaded microneedle patches for effectively regenerating skin tissues in UV-aging induced mice. Dissolving microneedle patches, composed of polyvinyl alcohol with an MW of 9–10 kDa and polyvinylpyrrolidone with an MW of 56 kDa, were fabricated using the double-casting method at three AMSC-MP concentrations: i.e., 30 % (MN30), 25 % (MN25), and 20 % (MN20). The microneedles patches were then evaluated for morphological, mechanical resistance, and insertion properties. An ex vivo release study was also conducted using the Franz cell method, and in vivo efficacy and irritation were then determined through collagen density scores, fibroblast cell counts, and skin irritation studies of UV-aging induced mice. The AMSC-MP microneedles displayed a pyramidal shape with 500 µm sharp tips. Mechanical testing revealed that MN30 achieved its deepest insertion into Parafilm® M (447.44 ± 37.21 µm), while MN25 achieved its deepest insertion into full-thickness porcine skin (717.92 ± 25.40 µm). The study revealed a controlled EGF release for up to 24 h, with MN20 exhibiting the highest deposition (55.94 ± 12.34 %). These findings demonstrate the successful penetration of microneedles through the stratum corneum and viable epidermis. Collagen density scores and fibroblast cell counts were significantly higher in all microneedle formulations than the control, with MN30 having the highest values. Inflammatory cell counts indicated minimal presence suggesting non-irritation in the in vivo study. Dissolving microneedle patches exhibited favorable characteristics and efficiently delivered AMSC-MP with minimal potential for irritation, providing potential technology for delivering biological anti-aging agents for the purposes of fostering skin regeneration.

Agent-based model demonstrates the impact of nonlinear, complex interactions between cytokinces on muscle regeneration.

Muscle regeneration is a complex process due to dynamic and multiscale biochemical and cellular interactions, making it difficult to identify microenvironmental conditions that are beneficial to muscle recovery from injury using experimental approaches alone. To understand the degree to which individual cellular behaviors impact endogenous mechanisms of muscle recovery, we developed an agent-based model (ABM) using the Cellular-Potts framework to simulate the dynamic microenvironment of a cross-section of murine skeletal muscle tissue. We referenced more than 100 published studies to define over 100 parameters and rules that dictate the behavior of muscle fibers, satellite stem cells (SSCs), fibroblasts, neutrophils, macrophages, microvessels, and lymphatic vessels, as well as their interactions with each other and the microenvironment. We utilized parameter density estimation to calibrate the model to temporal biological datasets describing cross-sectional area (CSA) recovery, SSC, and fibroblast cell counts at multiple timepoints following injury. The calibrated model was validated by comparison of other model outputs (macrophage, neutrophil, and capillaries counts) to experimental observations. Predictions for eight model perturbations that varied cell or cytokine input conditions were compared to published experimental studies to validate model predictive capabilities. We used Latin hypercube sampling and partial rank correlation coefficient to identify in silico perturbations of cytokine diffusion coefficients and decay rates to enhance CSA recovery. This analysis suggests that combined alterations of specific cytokine decay and diffusion parameters result in greater fibroblast and SSC proliferation compared to individual perturbations with a 13% increase in CSA recovery compared to unaltered regeneration at 28 days. These results enable guided development of therapeutic strategies that similarly alter muscle physiology (i.e. converting extracellular matrix [ECM]-bound cytokines into freely diffusible forms as studied in cancer therapeutics or delivery of exogenous cytokines) during regeneration to enhance muscle recovery after injury.

Agent-based model demonstrates the impact of nonlinear, complex interactions between cytokines on muscle regeneration

Muscle regeneration is a complex process due to dynamic and multiscale biochemical and cellular interactions, making it difficult to identify microenvironmental conditions that are beneficial to muscle recovery from injury using experimental approaches alone. To understand the degree to which individual cellular behaviors impact endogenous mechanisms of muscle recovery, we developed an agent-based model (ABM) using the Cellular-Potts framework to simulate the dynamic microenvironment of a cross-section of murine skeletal muscle tissue. We referenced more than 100 published studies to define over 100 parameters and rules that dictate the behavior of muscle fibers, satellite stem cells (SSCs), fibroblasts, neutrophils, macrophages, microvessels, and lymphatic vessels, as well as their interactions with each other and the microenvironment. We utilized parameter density estimation to calibrate the model to temporal biological datasets describing cross-sectional area (CSA) recovery, SSC, and fibroblast cell counts at multiple timepoints following injury. The calibrated model was validated by comparison of other model outputs (macrophage, neutrophil, and capillaries counts) to experimental observations. Predictions for eight model perturbations that varied cell or cytokine input conditions were compared to published experimental studies to validate model predictive capabilities. We used Latin hypercube sampling and partial rank correlation coefficient to identify in silico perturbations of cytokine diffusion coefficients and decay rates to enhance CSA recovery. This analysis suggests that combined alterations of specific cytokine decay and diffusion parameters result in greater fibroblast and SSC proliferation compared to individual perturbations with a 13% increase in CSA recovery compared to unaltered regeneration at 28 days. These results enable guided development of therapeutic strategies that similarly alter muscle physiology (i.e. converting extracellular matrix [ECM]-bound cytokines into freely diffusible forms as studied in cancer therapeutics or delivery of exogenous cytokines) during regeneration to enhance muscle recovery after injury.

POTENCY OF OCIMUM SANCTUM L EXTRACT OINTMENT ON THE MANAGEMENT PARTIAL-THICKNESS PALPEBRAL LACERATION, IS IT A NOVELTY TO BE CONSIDERED AS AN ALTERNATIVE THERAPY?: REVIEW ON HISTOPATHOLOGICAL CHANGES IN EXPERIMENTAL MODEL

Introduction and Objective: Ocimum sanctum extract (OSE) has been shown to have antioxidant, anti-inflammatory, and antibacterial properties which are responsible for wound healing. This study aims to analyze the effect of 5%, 10%, and 15% OSE ointment on changes in collagen density and fibroblast count in Wistar rats with partial-thickness palpebral laceration. Methods: Laboratory experimental study with randomized post-test only control group design. The subjects were 30 rats divided into 5 groups: The placebo; Gentamicin 0.3%; OSE 5%; OSE 10% and OSE 15% group. The intervention was administered 2 times/day for 4 days. On the fifth day, the palpebral tissue was obtained to make histopathological preparations and to observe the collagen density and fibroblast count. Data were analyzed using Mann-Whitney, One-Way ANOVA, Kruskal Wallis, and Tamhane test. A p-value <0.05 was assigned to the statistically significant. Results: The placebo group revealed loose collagen tissue and OSE 15% and gentamicin group revealed dense collagen tissue. The low fibroblast count was in the gentamicin and OSE 15% group and the highest fibroblast count was in the placebo group with a mean of 19.77±3.29; 22,67±0,73 and 60.9±10.66 cells, respectively. There was a significant difference (p<0.001) in the collagen density and the fibroblast count based on the treatment groups. Posthoc tests revealed differences in the collagen density level and the fibroblast count between the placebo and the whole group. Conclusion: There was a significant difference between placebo and all groups of collagen density changes and fibroblast count. There was no difference between OSE 15% and gentamicin both microscopically and statistically. Keywords: Ocimum sanctum L, Extract of Ocimum sanctum L, partial-thickness palpebral laceration, fibroblast cell count, collagen density

Effect of extract gel of Sargassum sp. against histopathological of the number of fibroblast cells in the burn healing process

Introduction: Burns are injuries caused by heat and cause damage to the skin. Burns are characterized by red, peeling skin. Burns must be treated properly so that wound healing can close completely. In Indonesia, seaweed is a promising commodity. From 5.2 million wet tons in 2011 to 9.2 million tons in 2013, seaweed output increased by 76.4 percent in Indonesia. This study aimed to study the histopathological picture of burn tissue given Sargassum sp. Methods: An experimental study design with a randomized post-test-only control group design was used for this study. White mice (Mus musculus), the study's population, made up this group. The study was divided into 4 treatment groups. The results of the calculation of the number of fibroblast cells were then evaluated using statistical tests to determine differences between groups. Data analysis was carried out using SPSS ver. 25 with a One-Way ANOVA test. Results: The mean result of fibroblast cell count in group 1 (plain gel) was 36.14 PLP, group 2 (Sargassum sp. 5%) was 51.28 PLP, group 3 (Sargassum sp. 10%) was 53.36 PLP and group 4 (Sargassum sp. bioplacenton) 20.05 PLP. The results of the statistical test showed that there was a significant difference between groups with a p-value of 0.03 (<0.05). Conclusion: The results of the histology show that there is a potential content of Sargassum sp. to affect wound healing.

Topical curcumin gel accelerates healing of II A – degree burns on male wistar rats (Rattus norvegicus)

Background: Burns can happen to anyone at any time, so they require wound care, which is essential to speed up wound healing. Curcumin gel is suspected to help this process, which is administered topically. Besides being cheap, easy to find, and minimal side effects, the content of curcumin helps a series of biological processes that accelerate the recovery of II A – degree burns, which have not been studied extensively recently. Aim: The study aims to prove that topical curcumin gel can accelerate the healing of II A-degree burns in male Wistar rats (Rattus norvegicus). Methods: This true experimental study with a post-test control group design involved 28 male Wistar rats (Rattus norvegicus), which were initially intervened by inducing a heat probe that stimulated the formation of an area of II A – degree burns. Wound profile analysis was conducted in the control and treatment groups within 7 and 14 days, respectively. After the wound biopsy, the significance of the parametric test results on the average velocity of epithelialization, number of fibroblasts, and collagen density was assessed. Results: Epithelialization rate (p<0.001), fibroblast cell count (p=0.007), and collagen density (p=0.011) after curcumin administration were higher than controls. The curcumin treatment group at 14 days showed the greatest significance among the other comparisons (p<0.05). The higher the speed of epithelialization, the higher the number of fibroblasts or collagen, respectively, in the control and treatment groups (r > 0.65; p <0.05). Conclusion: Topical administration of curcumin gel has the potential to be a hope for future remedies that initiate the improvement of the healing process of II A – degree burns properly.

Effect of autologous conditioned serum (ACS) on histological characteristics and expression of soft tissue IL-1β gene after horizontal ridge augmentation surgery.

Horizontal ridge augmentation surgeries are common procedures in periodontics. Histological changes in soft tissues affect the success of surgery in many ways. Autologous conditioned serum (ACS) increases interleukin-1 receptor antagonist (IL-1Ra) and causes inflammation modulation. Therefore, the present study aimed to investigate the effect of ACS on histological changes and gene expression of soft tissues after horizontal ridge augmentation surgeries. This double-blind split-mouth clinical trial was performed on patients needing implants with horizontal ridge augmentation (n=21). The control and intervention groups were considered split-mouth in two areas of the patients' oral soft tissues. A collagen membrane impregnated with ACS was used on the test side, and only a collagen membrane was used on the control side. After four months, histological changes in soft tissues, such as the amount of connective tissue collagen, fibroblast and inflammatory cell counts, and expression of the IL-1β gene, were evaluated. The fibroblast counts in the ACS group were significantly higher than in the control group. In addition, ACS caused a significant increase in the amount of collagen in the soft tissues compared to the control group (P<0.01). However, the number of inflammatory cells was similar in the two groups (P>0.05). IL-1β gene expression was not significantly different between the case and control groups. Under the limitations of the present study and based on the results of histological examinations, ACS increased the number of fibroblasts and the amount of collagen in soft tissues without affecting inflammatory cells (P=0.562).

“Ultraviolet C phototherapy as an adjunct wound therapy in full-thickness wound of a rat model”

Ultraviolet C (UVC) phototherapy is a potential modality to improve wound healing due to its well-known antimicrobial properties, and it promotes the expression of certain growth factors. However, limited data exist to show the clinical effect of UVC in wound healing compared with other advanced modern dressings. This animal preclinical study involved 56 Sprague–Dawley female rats aged 3 months old, weighing between 250 and 300 g, which were acclimatized for one week. Following the creation of a 2 × 2-cm-square full-thickness wound over the dorsum of each rat, they were divided into four treatment groups, namely, the control, UVC, hydrofiber silver, and UVC/hydrofiber silver groups. On Days 2, 4, 7, 10, 14, 21, and 28 postwounding, two rats from each group underwent wound assessment via wound measurement (mm2), calculation of the percentage of wound contracture and percentage of epithelization. Wound specimens were obtained for histological examination of inflammatory cells (neutrophils, lymphocytes and macrophages) and fibroblast cell counts. There was a relationship between wound size reduction and time to heal (P<0.05, R2=0.70) among the four treatment groups. The UVC/hydrofiber silver group had a significantly smaller wound size given the time to heal compared with the control group (P = 0.01) and UVC group (P = 0.02). There were no significant differences in terms of wound contracture and epithelization percentage among the four treatment groups. Histopathological examination revealed a significantly lower mean fibroblast count in the UVC/hydrofiber silver group than in the other groups (P = 0.025). These data suggested that UVC phototherapy did not increase the rate of healing but maintained the integrity of the wound by providing antimicrobial properties and preventing overproduction of fibroblasts. UVC was also safe, as no overt inflammatory response was discovered.

The Matrix Metalloproteinase Inhibitor Batimastat Reduces Epidural Fibrosis After Laminectomy in Rats.

To investigate the efficacy of locally applied batimastat after laminectomy in preventing postoperative epidural fibrosis. Thirty-two Wistar albino male rats weighing 200?250 g were used. The rats were assigned to four different groups (I-Control group, II-sham group, III-Laminectomy+Batimastat group, and IV-Laminectomy+SpongostanTM group). The rats were euthanized 28 days after surgery before TNF-?, IL6, IL-1?, IL10, TGF-?1, and MMP9 gene expression levels of tissue in the surgical area were determined with qPCR. TNF-?, IL6, and IL10 protein levels were also measured in both tissue and plasma. In addition, the surgical area was evaluated by histopathological and immunohistochemical methods. TNF-?, IL6, and IL-1? gene expression levels were higher in the batimastat group than in the control group. Whereas IL10 gene expression levels increased about two-fold in the sham and SpongostanTM groups, in the batimastat group, it was similar to that in the control group. TGF-?1 gene expression was three-fold higher in the sham group but was similar to that in the control group in both batimastat and SpongostanTM groups. MMP9 gene expression levels significantly decreased only in the batimastat group. In addition, fibrosis score, fibroblast cell count, inflammatory cell count, and CD105 expression decreased in the batimastat group relative to the control. Molecular and pathological examination results suggested that batimastat is an effective agent in reducing the occurrence of epidural fibrosis after laminectomy.

FC 102PD INDUCED ARTERIOLAR AND PERITONEAL PATHOMECHANISMS ARE PARTIALLY REVERSED AFTER KIDNEY TRANSPLANTATION

Abstract Background and Aims Due to the unphysiological composition of PD fluids, chronic peritoneal dialysis (PD) induces progressive peritoneal fibrosis, hypervascularization, and vasculopathy. The evolution of the PD membrane and vasculopathy following kidney transplantation (KTx) is largely unknown. Method Arteriolar and peritoneal tissues were obtained from 107 children with chronic kidney disease (CKD5), 72 children on PD (treated with neutral pH PD fluids, with low glucose degradation product content, GDP) and 21 children, who underwent KTx 4-5 weeks after a median 21 months of PD. Specimen underwent standardized digital quantitative histomorphometry. Molecular mechanisms were studied in omental arterioles microdissected from surrounding fat by multi-omics followed by Gene Set Enrichment Analysis (GSEA); key findings were validated in parietal tissues of independent, matched cohorts by quantitative immunohistochemistry (n=15/group). Results Arteriolar transcriptome and proteome GSEA revealed suppression of leucocyte migration and T-cell activation / secretory pathways regulation, of sprouting angiogenesis biological processes and of epithelial proliferation and cell cycle after KTx as compared to PD. Lipid / fatty acid metabolism, autophagy and ATP synthesis pathways were activated. Transcriptome analysis including KTx, PD and CKD5 specifically attributed regulation of arteriolar lipid and fatty acid metabolism to transplantation and comprised 140 transcripts; their regulation was confirmed on the proteome level. Hub gene fatty acid synthase was identified by protein interaction analysis (string-db.org). 15 arteriolar genes activated by PD were inactivated after KTx and included glucose metabolisms and cytoskeleton related transcripts. 24 transcripts and 10 corresponding proteins induced by PD were still active after KTx and associated with biological processes related to TGF-ß signaling, fibrosis and mineral absorption. In line with arteriolar multi-omics findings, peritoneal hypervascularization induced by chronic PD was reversed after Tx to CKD5 level. CD45 positive tissue infiltrating leucocytes count was reduced by 40% and was independently associated with microvessel density in multivariable analysis including PD vintage, daily GDP exposure and recent KTx. Peritoneal lymphatic vessel density, submesothelial thickness, activated fibroblast, fibrin deposit, macrophage and EMT cell counts remained unchanged after KTx compared to PD. Arteriolar lumen to vessel ratios (a marker of vasculopathy) were similar in both groups. Vessel-homeostasis-related proteins in independent, matched cohorts demonstrated increased caspase-3 abundance in peritoneal arterioles after KTx. Arteriolar VEGF-A, thrombospondin, angiopoietin1/2, and hypoxia-inducible factor-1 (HIF-1a) were unchanged, while submesothelial HIF-1a and angiopoietin1/2 were decreased after Tx, favoring vessel maturation. The abundance of the key driver of fibrosis, TGF-ß-effector pSMAD2/3, was unchanged in the peritoneum and arterioles after Tx. Conclusion Our multi-omics analyses of fat covered omental arterioles, not directly exposed to PD fluids, demonstrate inhibition of PD induced immune response and angiogenesis pathways, of glucose metabolism and cytoskeleton regulation to levels similar as seen in children with CKD5. Arteriolar lipid and fatty acid metabolism is selectively altered after KTx. Reversal of low GDP PD induced hypervascularization and inflammation of the parietal peritoneum after KTx, mirror molecular changes in omental arterioles, while profibrotic activity persists after KTx in omental arterioles and in the parietal peritoneum.

Detection of nuclear receptors in gingival samples of diabetic and nondiabetic periodontitis patients.

Diabetes and periodontitis are two chronic inflammatory diseases sharing specific etiopathogenetic mechanisms, and both cause severe inflammation and destruction. The present study aimed to determine the receptor expressions of peroxisome proliferative-activated receptor (PPAR)-γ, retinoid X receptor (RXR)-α, vitamin D receptor (VDR), and nuclear factor kappa B (NF-κB) expressions in healthy gingiva and diseased gingival samples with or without diabetes. Forty-five participants as (1) healthy controls (C), (2) periodontitis group (P), and (3) diabetes and periodontitis group (DP) were enrolled. Plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment levels (CAL), and bleeding on probing (BOP) were recorded in all participants. Two gingival biopsies from each participant were obtained, and one underwent histological tissue processing while the other underwent qRT-PCR analysis of nuclear receptors. Inflammatory and fibroblast cell counts, PPAR-γ, RXR-α, VDR, and NF-κB were evaluated. Fibroblast cells were lowest in the DP group and highest in the healthy group. PPAR-γ, VDR, RXR, and NF-κB expressions were higher in the healthy controls in the qRT-PCR analysis and similar in the other groups. Immunohistochemistry analysis also showed similar results. qRT-PCR results concluded that healthy gingival samples had higher PPAR-γ, RXR, VDR, and NF-κB expressions, and immunohistochemistry findings supported the results. In addition, healthy gingiva contained higher fibroblast cells and lower inflammatory cells.

Formulation and Evaluation of &lt;i&gt;Binahong &lt;/i&gt;Leaves Extract Gel on Wound Healing of the Palatal Mucosa

Wound healing is a complex and integrated process that requires the collective roles of various cells and tissues of the palatal mucosa. During the proliferation phase, the sum of fibroblasts increases in response to damaged oral tissues. Thus, the number of fibroblast cells in the palatal mucosa is a substantial indicator of wound healing. Binahong leaves possess the potential to accelerate wound healing by stimulating fibroblast proliferation. This study aimed to formulate and evaluate the effect of Binahong leaves extract (BLE) gel on wound healing of the palatal mucosa by investigating the fibroblast cell count in Wistar rats. This experimental study was carried out in several stages which included the collection and processing of fresh Binahong leaves followed by extraction with 70% ethanol using maceration method. The extract was formulated to be a gel product with a concentration of 3%, 5%, and 7%. BLE gel was administered to Wistar rats which were deliberately injured at the palatal mucosa by excision. Wound healing was assessed using hematoxylin-eosin staining and the number of fibroblast cells was counted. The result of fibroblast proliferation was analyzed by One Way ANOVA followed by Bonferroni’s test with a significance level of 95%. There were significant differences in fibroblast cell proliferation between all test groups except between base gel and Aloclair® gel, base gel and BLE gel 7%, Aloclair® gel and BLE gel 7%, BLE gel 3% and BLE gel 5%. Application of BLE gel 3% promotes better wound healing of the palatal mucosa of Wistar rats.

Methyl palmitate reversed estradiol benzoate-induced endometrial hyperplasia in female rats

Early detection and treatment of endometrial hyperplasia (EH) is mandatory for endometrial cancer prevention. Several bioactive agents of plant origin have been shown to elicit their chemotherapeutic effect against tumors and cancer via induction of mitochondrial permeability transition(mPT) pore opening. This research was therefore aimed at evaluating the potential chemopreventive effect of methyl palmitate (MP), on estradiol benzoate(EB)-induced EH, looking at the mitochondrial-mediated pathway and other possible mechanisms of action. Mitochondria were isolated using differential centrifugation. The mPT pore, mitochondrial ATPase (mATPase) activity, lipid peroxidation and cytochrome c release were determined by standard methods using spectrophotometer. Uterine interleukin 1b, MDA levels and SOD, GSH activities, were determined using commercially available kits. The uterine histological and immunohistochemical assessment of estrogen receptor (ERα), IL-1b and caspas-3 were carried out. The fibroblast cell count density was determined using histomorphometry. At all the concentrations of MP used, there was no significant induction of mPT pore opening, neither any enhancement of mATPase activity nor release of cytochrome c when compared to the control. Similar pattern of results were recorded for the in vivo study. However, there was marked increase in the uterine MDA and interleukin 1b levels, with concurrent decrease in SOD and GSH activities, in the EB-treated group, which was significantly reversed by MP co-administration. Endometrial Hyperplasia observed in the EB-treated group was ameliorated by MP co-administration. The immunoexpression of ERα and IL-1b in the EB-treated group was reversed by MP co-administration. This study suggests anti-inflammatory, antioxidant and anti-proliferative potential of MP against EB-induced EH.

The effects of human amniotic membrane on silicone related capsule formation in rats

Silicone breast implants are commonly used materials in plastic surgery for breast augmentation and reconstruction and the most severe complication of silicone implants are capsule contraction which occurs in 40% of patients. The aim of our study is to evaluate how the amniotic membrane alters the capsule formation effects of silicone 24 wistar rats were used in the study. We placed a bare silicone block into the left side (Subgroup A) and single layer amniotic membrane coated silicone block into the right side (Subgroup B) of the rats back. The rats were then separated into three groups and in group 1 rats were euthanized after 3 weeks, in group 2 after 12 weeks and in group 3 after 24 weeks. Then capsule thickness, fibroblast and lymphocyte cell counts were evaluated for each sample. In Group 2 and group 3, the capsule thickness in Subgroup B was detected to be statistically significantly lower than that in Subgroup A. In Group 1, 2, and 3, the lymphocyte count in the capsule tissue taken from Subgroup B was lower than Subgroup A but the difference was not statistically significant. In Group 2 and 3, the fibrocyte count detected in the capsule tissue in Subgroup B was found to be statistically significantly lower than Subgroup A. the amniotic membrane was demonstrated to reduce capsule thickness by the antifibrinolytic effect in our study.

Evaluation of the Combination of Sargassum Duplicatum, Sargassum Ilicifolium, Abelmoschus Esculentus, and Garcinia Mangostana Extracts for Open Wound Healing in Diabetic Mice

Introduction: Diabetes mellitus (DM) is a chronic disease with diverse and complex metabolic disorders. DM can be identified with hyperglycemic conditions. In 2017, Indonesia ranked sixth in the world for the highest number of diabetic patients after China, India, United States, Brazil, and Mexico. Objective: This study aims to determine the combined effect of alginate extract from Sargassum duplicatum and Sargassum ilicifolium with okra (Abelmoschus esculentus) and mangosteen (Garcinia mangostana) pericarp extracts to ameliorate the wound width process, the number of neutrophil cells, macrophages, fibroblasts, fibrocytes, and collagen density in streptozotocin-induced diabetic mice. Methods: This study used male mice (strain BALB/C, weighting 30±5 g each) which were grouped into four groups; normal control with water only; diabetic control group; combination treatment group with S. duplicatum extract, A. esculentus extract, and G. mangostana extract; and combination treatment group with S. ilicifolium extract, A. esculentus extract, and G. mangostana extract. The diabetic control group and combination treatment groups were injected with multiple low doses of streptozotocin (STZ) at 30 mg/kg BW intraperitoneally. The combination extract dose given in treatment groups was 50 mg/kg BW. Measurement of fasting blood sugar levels was done before and after the STZ injection. The mice’s skin was cut in the gluteal section as long as 1 cm, then treated topically in three different time periods namely 3, 7, and 14 days. Histological preparation and analysis were conducted on the 15th day. Results: The administration of A. esculentus extract and G. mangostana extract combined with S. duplicatum and S. ilicifolium increased wound healing indicator rate, decreased neutrophile cell count (NCC) and fibrocyte cell count (FCCC), and increased collagen synthesis as an extracellular matrix (ECM). The treatments could restore the open wound healing process of diabetic mice to their normal state. There were no significant differences in macrophage cell count (MCC) and fibroblast cell count (FBCC). The histopathology showed that the combination treatment groups also could accelerate the wound healing process in mice skin. Conclusion: In summary, this study addressed that the administration of A. esculentus extract and G. mangostana extract combined with S. duplicatum and S. silicifolium extracts could ameliorate open wound healing in diabetic mice.

Effects of Decompressive Craniectomy with Mesh in the Level of Transforming Growth Factor Beta, Fibroblast Cell Count, Size of Fibrosis, and Clinical Adhesion of Peridural Tissues in Wistar Rat with Traumatic Brain Injury

One option to control intracranial pressure (ICP) is by performing decompressive craniectomy (DC) before definitive treatment. A common problem after DC is adhesion. Separated injured tissue surfaces by using bioabsorbable membranes over a critical/certain period are predicted to prevent fibrin bridge formation and adhesion. This study is conducted to prove the effects of DC with mesh on TGF-β levels, fibroblast cell count, fibrosis size and clinical adhesions in wistar rat with traumatic brain injury. The current research uses animal model with randomized post-test only control group design. A total of 20 samples that met the eligibility criteria were included in the study and randomized. A total of 10 samples were grouped as a control group (standard DC) and 10 samples were grouped as a study group (DC with mesh). On day 7, we performed euthanasia and took peridural tissue for examination of TGF-β levels, fibroblast cell count, and fibrosis size. The mean results of TGF-β in the study group were lower than the control group (81.28±17.48 pg/dl vs 93.83±9.51 pg/dl) although this difference was not statistically significant (p=0.062). For the mean number of fibroblasts cells, there was a significant difference where the study group was lower than the control group (56.8±20.59 cells/HPF vs 94.8±18.56 cells/HPF) with p&lt;0.001. The mean size of peridural fibrosis also showed a significant difference where the mean size of peridural fibrosis in the study group was lower than the control group (1182.52±347.05 μm vs 1545.78±378.28 μm) with p=0.038. Only DC samples (without the use of mesh) had a relative risk 8 times greater for clinical adhesions compared to mesh (RR=8; 95% CI, 1,215-52,693; p=0,005). DC with mesh significantly resulted in low fibroblast cells count, size of peridural fibrosis, and the risk of clinical adhesions in wistar rat with traumatic brain injury.

Evaluation of apoptosis and hypoxia-related factors in gingival tissues of smoker and non-smoker periodontitis patients.

Smoking causes pathological changes in all tissues, including gingiva and alveolar bone. The aim of present study was to evaluate apoptotic tissue alterations and tissue destruction in smoker and non-smoker periodontitis patients and healthy individuals. Gingival biopsy samples from 15 systemically and orally healthy individuals (Group 1), 15 systemically healthy periodontitis patients (Group 2), 15 systemically and orally healthy smokers (Group 3), and 15 systemically healthy smoker periodontitis patients (Group 4) were enrolled in the present study. Clinical periodontal measurements as plaque index (PI), gingival index (GI), and clinical attachment levels (CAL) were recorded, and gingival biopsies were obtained. Biopsy samples were fixed in formalin solution and embedded in paraffin. Fibroblast and inflammatory cell counts were determined via histomorphometrically. Hypoxia-inducible factor alpha (HIF-1α), vascular endothelial growth factor(VEGF), tissue inhibitor of matrix metalloproteinase-1(TIMP-1), matrix metalloproteinases-8(MMP-8) expressions, Bax, Bcl-2, and caspase-3 expressions were evaluated via immunohistochemistry. Demographic data of the study groups were similar. Smoking levels of the smokers were also similar. The highest fibroblast cell counts were observed in healthy controls and the counts were similar in other groups. The highest inflammatory cell counts were found in smoker periodontitis group, and the lowest counts were found in healthy control groups. The differences were statistically significant. HIF-1α and Bax expressions were elevated and Bcl-2 decreased in smoker periodontitis patients compared with healthy individuals. However, there were no differences in VEGF, MMP-8, and TIMP-1 expressions. Within limits of present study, it can be suggested that both smoking and periodontitis caused similar decrease in fibroblast counts while causing a dramatic increase in inflammatory cell counts. Increased apoptosis and hypoxia also accompanied to the increased inflammation.

Protective effects of alpha stone on monosodium glutamate-induced uterine hyperplasia in female wistar rats

BackgroundUterine leiomyomas (fibroids), a menace of the reproductive age, is characterized by proliferation of smooth muscle cells (hyperplasia) of the uterus. Alpha Stone Decoction is a poly-herbal formulation that is used for the shrinkage and prevention of uterine fibroids in folkore medicine. ObjectiveWe investigated the efficacy and safety of Alpha Stone Decoction (ASD), on monosodium glutamate (MSG) induced uterine hyperplasia. Materials and methodsTwenty-eight mature virgin female rats were randomly divided into four study groups: A-control B- MSG (200 mg/kgbw), C- MSG + ASD (100 mg/kgbw) and D- ASD 100 mg/kgbw alone. The administration was carried out by as a single daily dose via intraperitoneal route for 14 days. Total protein, triglycerides, estradiol (estrogen), progesterone, and total cholesterol levels in sera were determined using appropriate kits. Uterine hyperplasia was assessed via histomorphometric method using the mitotic image plus software to compute the fibroblast cell count density while the uteri and ovaries of animals were stained with mason-tricon stain for histological examination. ResultsAdministration of MSG for 14 days resulted heavy deposits of collagen connective tissue within the myometrium layers of the uteri. ASD significantly (p < 0.05) reduced fibroblast cell count in MSG-treated animals and also protected against MSG-induced damage observed in the myometrium of the uteri and ovaries of the animals. Significant increases (p < 0.05) in levels of total protein; triglycerides, progesterone, cholesterol and estrogen in the MSG-treated animals were ameliorated following administration of ASD. ConclusionThese findings suggest that ASD contains bioactive agents which reversed MSG-induced uterine hyperplasia. It may therefore be useful in reducing the proliferation of fibroblast cells and managing other symptoms associated with uterine myoma.

Histological evaluation of peri-implant mucosal and gingival tissues in peri-implantitis, peri-implant mucositis and periodontitis patients: a cross-sectional clinical study

Objective: Aim of present study was to evaluate gingival tissue samples obtained from healthy and diseased sites of teeth and dental implants in terms of hypoxia and collagenase activity.Methods: Four study groups were created as Group-1; healthy individuals (H), Group-2; periodontitis patients with stage 3 grade B (P), Group-3; patients with peri-implant mucositis. Group-4; patients with peri-implantitis (P-IMP). Plaque index (PI), gingival index (GI) and probing pocket depth (PPD) were recorded. Gingival and peri-implant mucosal biopsies were obtained. Fibroblast and inflammatory cells were counted. Hypoxia-inducible factor (HIF)-1α, prolyl hydroxylase (PH), matrix metalloproteinase (MMP)-8, tissue inhibitor of MMPs (TIMP)-1, cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) levels were determined via immunohistochemistry.Results: Healthy controls had highest fibroblast cell counts and lowest inflammatory cell counts compared to other groups. Peri-implantitis and periodontitis samples had similar fibroblast and inflammatory cell counts, while peri-implant mucositis had higher fibroblast cells and lowered inflammatory cells compared to periodontitis and peri-implantitis samples. HIF-1α, COX-2 and iNOS levels were lowest in healthy controls and increased in other groups. Peri-implant mucositis samples had significantly lower expressions of HIF-1α, COX-2 and iNOS compared to peri-implantitis and periodontitis groups. PH expressions were lower in periodontitis and peri-implantitis groups compared to healthy controls and peri-implant mucositis groups. MMP-8 levels were lower in healthy group compared to other groups while levels were similar in periodontitis, peri-implant mucositis and peri-implantitis groups. TIMP levels were similar in groups.Conclusion: Periodontitis, peri-implantitis, and peri-implant mucositis samples exhibited higher inflammation and lower fibroblast cell counts and tend to have increased tissue collagenase activity, hypoxia and inflammation compared to healthy samples.

Hypoxia and collagen crosslinking in the healthy and affected sites of periodontitis patients

Summary: Present study suggests that diseased sites of periodontitis with stage 3 grade B and C had decreased fibroblast cell density, hypoxia-inducible factor (HIF) and vascular endothelial growth factor (VEGF) expressions while increased inflammatory cell counts compared to both healthy sites of the periodontitis patients and healthy controls. Collagen maturation enzymes also decreased in the diseased sites.Objective: The present study aimed at determining markers of hypoxia and collagen crosslinking in healthy and diseased gingiva from healthy individuals and periodontitis patients.Methods: Group-1; healthy individuals, Group-2; healthy sites of periodontitis patients-stage 3 grade B, (H-GradeB) Group-3; diseased sites of periodontitis patients-stage 3 grade B, (D-GradeB). Group-4; healthy sites of periodontitis patients-stage 3 grade C, (H-GradeC). Group-5; diseased sites of periodontitis patients-stage 3 grade C, (D-GradeC). Plaque index (PI), gingival index (GI) and clinical attachment levels (CALs) were recorded. Gingival biopsies were obtained. Fibroblast and inflammatory cells were counted. HIF-1α, prolyl hydroxylase (PH), VEGF, lysyl oxidase (LOX) and lysyl hydroxylase (LH) levels were determined via immunohistochemistry.Results: Fibroblast cell counts were lower in D-GradeC and D-GradeB than other groups. C group had highest fibroblast cell counts. Inflammatory cell counts were highest in the D-GradeC and lowest in C group. HIF-1α levels were highest in C group and decreased in diseased sites. Lowest value was observed in D-GradeC group. VEGF, PH, and LH levels were higher in the control group compared to other groups. LOX levels were similar in the groups except for D-GradeC. LOX levels were similar in the groups except for D-GradeC which is significantly lower than those of the control group and healthy sites.Conclusions: The results revealed that diseased sites of periodontitis patients had decreased fibroblast cells, HIF and VEGF expressions while increased inflammatory cells. Collagen crosslinking tend to decrease with disease regardless of stage and grade of disease.