Osteoblast Growth Research Articles

Effect of amoxicillin and clindamycin on the gene expression of markers involved in osteoblast physiology

Background/purposeAmoxicillin and clindamycin are the most effective decontaminants for intraoral bone grafts before their application in bone regeneration without cytotoxic effects on osteoblasts, but their effects on the gene expression of markers involved in osteoblast growth and differentiation remain unclear. The study objective was to determine the effects of amoxicillin and clindamycin on the gene expression of markers involved in osteoblast growth and differentiation. Materials and methodsReal-time polymerase chain reaction (RT-PCR) was performed to explore the effect of 150 μg/mL clindamycin or 400 μg/mL amoxicillin on the gene expression by primary human osteoblasts (HOBs) of runt-related transcription factor 2 (Runx-2), osterix (OSX), alkaline phosphatase (ALP), osteocalcin (OSC), osteoprotegerin (OPG), receptor activator for nuclear factor κ B ligand (RANKL), type I collagen (Col-I), bone morphogenetic proteins 2 and 7 (BMP-2 and BMP-7), TGF-β1 and TGF-β receptors (TGF-βR1, TGF-βR2, and TGF-βR3), and vascular endothelial growth factor (VEGF). ResultsTreatment with 150 μg/mL clindamycin significantly increased the gene expression of TFG-β1, TGF-βR1, TGF-βR2, TGF-βR3, RUNX-2, Col-1, OSX, OSC, BMP-2, BMP-7, ALP, VEGF, and RANKL by HOBs. Treatment with 400 μg/mL amoxicillin significantly increased the gene expression of TGF-β R1, Col-I, OSC, RANKL, and OPG alone. ConclusionThese findings suggest that 150 μg/mL clindamycin is the decontaminant of choice to treat intraoral bone grafts before their application in bone regeneration. The osteogenic and antibacterial properties of clindamycin can favor and accelerate the integration of bone grafts in the oral cavity.

Preparation and characterization of different micro/nano structures on the surface of bredigite scaffolds

The preparation of controllable micro/nano structures on the surface of the bredigite scaffold is expected to exhibit the same support and osteoconductive capabilities as living bone. However, the hydrophobicity of the white calciμm silicate scaffold surface restricts the adhesion and spreading of osteoblasts. Furthermore, during the degradation process of the bredigite scaffold, the release of Ca2+ results in an alkaline environment around the scaffold, which inhibits the growth of osteoblasts. In this study, the three-dimensional geometry of the Primitive surface in the three-periodic minimal surface with an average curvature of 0 was used as the basis for the scaffold unit cell, and a white hydroxyapatite scaffold was fabricated via photopolymerization-based 3D printing. Nanoparticles, microparticles, and micro-sheet structures with thicknesses of 6 μm, 24 μm, and 42 μm, respectively, were prepared on the surface of the porous scaffold through a hydrothermal reaction. The results of the study indicate that the micro/nano surface did not affect the morphology and mineralization ability of the macroporous scaffold. However, the transition from hydrophobic to hydrophilic resulted in a rougher surface and an increase in compressive strength from 45 to 59–86 MPa, while the adhesion of the micro/nano structures enhanced the scaffold's ductility. In addition, after 8 days of degradation, the pH of the degradation solution decreased from 8.6 to around 7.6, which is more suitable for cell growth in the hμman body. However, there were issues of slow degradation and high P element concentration in the degradation solution for the microscale layer group during the degradation process, so the nanoparticle and microparticle group scaffolds could provide effective support and a suitable environment for bone tissue repair.

The histone acetyltransferase Mof regulates Runx2 and Osterix for osteoblast differentiation.

Osteoblast differentiation is regulated by various transcription factors, signaling molecules, and posttranslational modifiers. The histone acetyltransferase Mof (Kat8) is involved in distinct physiological processes. However, the exact role of Mof in osteoblast differentiation and growth remains unknown. Herein, we demonstrated that Mof expression with histone H4K16 acetylation increased during osteoblast differentiation. Inhibition of Mof by siRNA knockdown or small molecule inhibitor, MG149 which is a potent histone acetyltransferase inhibitor, reduced the expression level and transactivation potential of osteogenic key markers, Runx2 and Osterix, thus inhibiting osteoblast differentiation. Besides, Mof overexpression also enhanced the protein levels of Runx2 and Osterix. Mof could directly bind the promoter region of Runx2/Osterix to potentiate their mRNA levels, possibly through Mof-mediated H4K16ac to facilitate the activation of transcriptional programs. Importantly, Mof physically interacts with Runx2/Osterix for the stimulation of osteoblast differentiation. Yet, Mof knockdown showed indistinguishable effect on cell proliferation or apoptosis in MSCs and preosteoblast cells. Taken together, our results uncover Mof functioning as a novel regulator of osteoblast differentiation via the promotional effects on Runx2/Osterix and rationalize Mof as a potential therapeutic target, like possible application of inhibitor MG149 for the treatment of osteosarcoma or developing specific Mof activator to ameliorate osteoporosis.

Myricetin exerts anti-biofilm activity and attenuates osteomyelitis by inhibiting the TLR2/MAPK pathway in experimental mice

AimsTo evaluate the potential of Myricetin against S.aureus induced osteomyelitis. BackgroundOsteomyelitis is infected condition of bone by micro-organisms. The mitogen-activated protein kinase (MAPK), inflammatory cytokines and Toll-like receptor-2 (TLR-2) pathway are mainly involved in osteomyelitis. Myricetin is a plant-food derived flavonoid which shows anti-inflammatory activity. ObjectiveIn the present study, we evaluated the potential of Myricetin against S.aureus induced osteomyelitis. MC3T3-E1 cells were used for in vitro studies. MethodMurine model of osteomyelitis was developed in BALB/c mice by injecting S.aureus in the medullary cavity of the femur. The mice were studied for bone destruction, anti-biofilm activity, osteoblast growth markers alkaline phosphatase (ALP), osteopontin (OCN) and collagen type-I (COLL-1) were studied by RT-PCR, ELISA analysis for levels of proinflammatory factors CRP, IL-6 and IL-1β. Expression of proteins by Western blot analysis and anti-biofilm effect by Sytox green dye fluorescence assay. Target confirmation was done by performing in silico docking analysis. ResultsMyricetin reduced bone destruction in osteomyelitis induced mice. The treatment decreased bone levels of ALP, OCN, COLL-1 and TLR2. Myricetin decreased serum levels of CRP, IL-6 and IL-1β. The treatment suppressed activation of MAPK pathway and showed anti-biofilm effect. Docking studies suggested high binding affinity of Myricetin with MAPK protein in silico, by showing lower binding energies. ConclusionMyricetin suppresses osteomyelitis by inhibiting ALP, OCN, COLL-1 via the TLR2 and MAPK pathway involving inhibition of biofilm formation. In silico studies suggested MAPK as potential binding protein for myricetin.

Design and Development of Tissue Engineering Materials based on Imine Bonds

Hyaluronic acid was oxidized to obtain hyaluronic acid containing aldehyde groups (ALD-HA). Then it was reacted with PETMP-AU, the photo-initiated reaction product of pentaerythritol tetra(3-mercaptopropionate) and allylurea, to synthesize a bioscaffold material with dual degradation behaviors, both to hyaluronidase and to weakly acidic environment. The material has a heterogeneous porous micromorphology with a pore size ranging from 10 to 100 μm. The growth of osteoblasts in the polymer was observed. The MC3T3-E1 osteoblasts encapsulated in the polymer could adhere to the inner wall of the pores and grow three-dimensionally. Moreover, the cells could continue to divide and proliferate and grow well. It shows that the material has good biocompatibility and can be used for biological tissue engineering materials.

Effects of Electronic Cigarette E-Liquids on the Adhesion and Growth of Osteoblast Cells

Electronic cigarettes (E-cigarettes) were designed to replace traditional cigarettes to decrease the harmful effects of smoking. The liquid in e-cigarettes contains different chemicals, such as Propylene Glycol (PG) and Vegetable Glycerin (VG), and may contain nicotine. To be more attractive, e-liquids are supplemented with flavorings. Because vaped e-liquids are shown to be damaging to oral tissues, similar effects could occur in the oral cavity with non-vaped e-liquids. We evaluated the effects of non-vaped e-liquid constituents on osteoblast behavior. For this purpose, human osteoblast cells (MG-63) were exposed to e-liquid containing 70% PG -30% VG with and without tobacco flavor and with or without nicotine at 12, 18 mg/mL, or no nicotine content. The e-liquids were used at various concentrations (0, 1, and 5%). To evaluate the effect of the e-liquids on osteoblast morphology and adhesion, optical microscope observations and viable cell counting using trypan blue exclusion were used. Cell growth was also analyzed using the Methyl Thiazol Tetrazolium (MTT) assay, while cytotoxicity was tested by measuring Lactate Dehydrogenase (LDH) levels after cell exposure for 24 h to the e-liquids. Our results show that e-liquids induced significant morphological changes evidenced by round cell forms, with no contact between cells. Adhesion was significantly reduced, particularly in the presence of nicotine. E-liquids at a concentration of 5% significantly reduced osteoblast growth. This effect was observed with both flavored and non-flavored e-liquid with or without nicotine. The decreased osteoblast adhesion and growth after exposure to e-liquid was confirmed by increased levels of LDH. Overall results indicate the potentially harmful effects of e-liquid non-vaped chemicals on bone cells, which could lead to the impairment of bone regeneration and tissue remodeling processes.

Yam-derived exosome-like nanovesicles stimulate osteoblast formation and prevent osteoporosis in mice.

Plants-releasing exosome-like nanovesicles (PENs) contain miRNA, bioactive lipids, mRNAs, and proteins to exert antioxidant, anti-inflammatory, and regenerative activity. Substances extracted from yams have been reported to promote osteoblast growth in bone regeneration, which prevent weak and brittle bones in osteoporosis. Herein, we describe the beneficial effects of yam-derived exosome-like nanovesicles (YNVs) on promoting differentiation and mineralization of osteoblasts for bone regeneration in ovariectomized (OVX)-induced osteoporotic mice. YNVs were successfully isolated and characterized. YNVs stimulate the proliferation, differentiation, and mineralization of osteoblasts with increased bone differentiation markers (OPN, ALP, and COLI). Interestingly, YNVs do not contain saponins including diosgenin and dioscin known to mainly exert osteogenic activity of yams. Instead, the osteogenic activity of YNVs was revealed to be resulted from activation of the BMP-2/p-p38-dependent Runx2 pathway. As a result, YNVs promote longitudinal bone growth and mineral density of the tibia in the OVX-induced osteoporotic mice in vivo, and these results positively correlate the significant increases in osteoblast-related parameters. In addition, the orally administered YNVs were transported through the GI tract and absorbed through the small intestine. These results showed an excellent systemic biosafety determined by histological analysis and liver/kidney toxicity tests. Taken together, YNVs can serve as a safe and orally effective agent in the treatment of osteoporosis.

In-Vitro Evaluation of Photofunctionalized Implant Surfaces in a High-Glucose Microenvironment Simulating Diabetics

The present study aimed to assess the efficacy of photofunctionalization on commercially available dental implant surfaces in a high-glucose environment. Discs of three commercially available implant surfaces were selected with various nano- and microstructural alterations (Group 1-laser-etched implant surface, Group 2-titanium-zirconium alloy surface, Group 3-air-abraded, large grit, acid-etched surface). They were subjected to photo-functionalization through UV irradiation for 60 and 90 min. X-ray photoelectron spectroscopy (XPS) was used to analyze the implant surface chemical composition before and after photo-functionalization. The growth and bioactivity of MG63 osteoblasts in the presence of photofunctionalized discs was assessed in cell culture medium containing elevated glucose concentration. The normal osteoblast morphology and spreading behavior were assessed under fluorescence and phase-contrast microscope. MTT (3-(4,5 Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and alizarin red assay were performed to assess the osteoblastic cell viability and mineralization efficiency. Following photofunctionalization, all three implant groups exhibited a reduced carbon content, conversion of Ti4+ to Ti3+, increased osteoblastic adhesion, viability, and increased mineralization. The best osteoblastic adhesion in the medium with increased glucose was seen in Group 3. Photofunctionalization altered the implant surface chemistry by reducing the surface carbon content, probably rendering the surfaces more hydrophilic and conducive for osteoblastic adherence and subsequent mineralization in high-glucose environment.

The effect of alginate scaffolds on bone healing in defects formed with drilling model in rat femur diaphysis.

Alginate (ALG) is a biocompatible and biodegradable polymer. Mechanical weakness is one of the main problems for the alginate-based scaffolds. Various plasticizer additives or modifications tested to improve the mechanical properties. In the presented study, ALG plasticized with triacetin (TA), and tributyl citrate (TBC) than tested on bone healing. In the presented study, the alginate modified with triacetin or tributyl citrate. In-vitro, and in-vivo efficiency of the scaffolds tested on bone tissue regeneration. Scaffolds fabricated by solvent casting, and physicochemical characterizations performed. Monocytes (THP-1) cultured with scaffolds, and macrophage-released cytokines was determined. In-vivo efficacy of the scaffolds was tested in the rat drill hole model. Alginate and tributyl citrate-modified scaffolds have no cytotoxic effect on osteoblastic cells (MC-3T3). Tributyl citrate modification increased tumor necrosis factor-alpha (TNF-alpha) level but did not increase interleukin -1 beta (IL-1 beta) level. In vivo studies showed that osteoblastic growth was significant in alginate and triacetin-modified scaffolds. However, the best values for osteoclastic activity and osteoid tissue formation seen in the triacetin modification. The results demonstrated that the modified alginate scaffolds were more successful than non-modified alginate scaffolds and can used as long-term bone repairing treatments.

The biological responses of osteoblasts on titanium: Effect of oxygen level and surface roughness

Due to the general application of invitro test, cell culture is generally selected to evaluate the cytocompatibility of devices and materials. The choice of test condition should depend on the probable site and clinical application. The oxygen content of human body could be estimated around 5%∼12%, and the oxygen level of healing bone fracture range from 0.8%∼3.8%%. However, materials for bone implant are traditionally evaluated under laboratory normoxia condition (21% O2) invitro. The aim was to study the effect of oxygen level on osteoblast upon high stiffness titanium with different roughness. After sandblasted and acid-etched (SLA) process, we create titanium surfaces with four different roughness. The differentiation and proliferation of MC3T3-E1 osteoblast cultured on SLA-treated specimens were evaluated in designed chamber with oxygen level of 1%, 5%, 10%, 21%. By scanning electron microscopy, all samples had sub-micro pit inside the micro-holes upon SLA-treated Ti disk surface. The decrease of oxygen level from 21% to 5% promoted osteoblast growth of SLA-treated specimens, but 1% O2 delayed cell proliferation. The surface roughness of specimens influenced osteoblast cell differentiation. The differentiation and proliferation ability of the cells upon SLA-treated specimens is proportional to oxygen level. Our results demonstrated that 5% O2 will easily discriminate osteoblasts responses on different SLA-treated specimens. These results suggest that hypoxia (5% O2) environment is better model for biological evaluation of bone-related materials.

3D Printed Hierarchical Porous Poly(ε-caprolactone) Scaffolds from Pickering High Internal Phase Emulsion Templating.

In the realm of biomaterials, particularly bone tissue engineering, there has been a great increase in interest in scaffolds with hierarchical porosity and customizable multifunctionality. Recently, the three-dimensional (3D) printing of biopolymer-based inks (solutions or emulsions) has gained high popularity for fabricating tissue engineering scaffolds, which optimally satisfies the desired properties and performances. Herein, therefore, we explore the fabrication of 3D printed hierarchical porous scaffolds of poly(ε-caprolactone) (PCL) using the water-in-oil (w/o) Pickering PCL high internal phase emulsions (HIPEs) as the ink in 3D printer. The Pickering PCL HIPEs stabilized using hydrophobically modified nanoclay comprised of aqueous poly(vinyl alcohol) (PVA) as the dispersed phase. Rheological measurements suggested the shear thinning behavior of Pickering HIPEs having a dispersed droplet diameter of 3-25 μm. The pore morphology resembling the natural extracellular matrix and the mechanical properties of scaffolds were customized by tuning the emulsion composition and 3D printing parameters. In vitro biomineralization and drug release studies proved the scaffolds' potential in developing the apatite-rich bioactive interphase and controlled drug delivery, respectively. During in vitro osteoblast (MG63) growth experiments for up to 7 days, good adhesion and proliferation on PCL scaffolds confirmed their cytocompatibility, assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) analysis. This study suggests that the assembly of HIPE templates and 3D printing is a promising approach to creating hierarchical porous scaffolds potentially suitable for bone tissue engineering and can be stretched to other biopolymers as well.

Nuclear receptor modulators inhibit osteosarcoma cell proliferation and tumour growth by regulating the mTOR signaling pathway

Osteosarcoma is the most common primary malignant bone tumour in children and adolescents. Chemoresistance leads to poor responses to conventional therapy in patients with osteosarcoma. The discovery of novel effective therapeutic targets and drugs is still the main focus of osteosarcoma research. Nuclear receptors (NRs) have shown substantial promise as novel therapeutic targets for various cancers. In the present study, we performed a drug screen using 29 chemicals that specifically target 17 NRs in several different human osteosarcoma and osteoblast cell lines. The retinoic acid receptor beta (RARb) antagonist LE135, peroxisome proliferator activated receptor gamma (PPARg) antagonist T0070907, liver X receptor (LXR) agonist T0901317 and Rev-Erba agonist SR9011 significantly inhibited the proliferation of malignant osteosarcoma cells (U2OS, HOS-MNNG and Saos-2 cells) but did not inhibit the growth of normal osteoblasts. The effects of these NR modulators on osteosarcoma cells occurred in a dose-dependent manner and were not observed in NR-knockout osteosarcoma cells. These NR modulators also significantly inhibited osteosarcoma growth in vivo and enhanced the antitumour effect of doxorubicin (DOX). Transcriptomic and immunoblotting results showed that these NR modulators may inhibit the growth of osteosarcoma cells by regulating the PI3K/AKT/mTOR and ERK/mTOR pathways. DDIT4, which blocks mTOR activation, was identified as one of the common downstream target genes of these NRs. DDIT4 knockout significantly attenuated the inhibitory effects of these NR modulators on osteosarcoma cell growth. Together, our results revealed that modulators of RARb, PPARg, LXRs and Rev-Erba inhibit osteosarcoma growth both in vitro and in vivo through the mTOR signaling pathway, suggesting that treatment with these NR modulators is a novel potential therapeutic strategy.

The overview of antimicrobial peptide‐coated implants against oral bacterial infections

AbstractDental implants are the most common therapeutic approach for resolving tooth loss and damage. Despite technical advances in treatment, implant failure rates can be as high as 23% with the major cause of peri‐implantitis: a multi‐species bacterial infection. With an annual growth rate in implant placements of 8.78% per annum, implant failure caused by bacterial infection is a significant oral and general health issue. The rise in antibiotic resistance in oral bacteria further adds pressure to implant failure; thus, there is a need for adjunctive therapy to improve implant outcomes. Due to the broad spectrum of activity and a low risk of inducing bacterial resistance, peptide antibiotics are emerging as a promising implant coating material to reduce/prevent peri‐implantitis and improve dental implant success rates. In this review, we summarised the current strategies of coating antimicrobial peptides (AMPs) onto dental implant material surfaces with multi‐functional properties to enhance osteoblast growth and prevent bacterial infections. This review compared the recent reported literature on dental implant coating with AMPs, which will provide an overview of the current dental implant coating strategies using AMPs and insights for future clinical applications.

Study on the treatment of postmenopausal osteoporosis with quercetin in Liuwei Dihuang Pill based on network pharmacology

BackgroundLiuwei Dihuang Pill (LP) was verified to alleviate postmenopausal osteoporosis (PMOP) development. Nevertheless, the major constituent of LP and the related network pharmacology study remain unexplored.MethodsProtein–protein interaction was established to identify the downstream target of LP in PMOP, and the related signaling pathway was investigated by bioinformatics analysis. MC3T3-E1 cells were added to ferric ammonium citrate (FAC) to mimic osteoporosis in vitro. The osteoblasts were identified by Alizarin red staining. Western blot was applied to evaluate protein levels. In addition, Cell Counting Kit-8 (CCK8) assay was applied to assess cell viability, and cell apoptosis was assessed by flow cytometry.ResultsQuercetin was the major constituent of LP. In addition, quercetin significantly reversed FAC-induced inhibition of osteogenic differentiation in MC3T3-E1 cells. In addition, quercetin notably abolished the FAC-induced upregulation of Bax, Caspase-3, FOS, JUN, TGFB1 and PPARD. In contrast, Bcl-2, p-mTOR/mTOR, p-AKT/AKT and p-PI3K/PI3K levels in MC3T3-E1 cells were reduced by FAC, which was restored by quercetin. Meanwhile, FAC notably inhibited the viability of MC3T3-E1 cells via inducing apoptosis, but this impact was abolished by quercetin. Furthermore, quercetin could reverse pcDNA3.1-FOS-mediated growth of FAC-treated osteoblasts by mediating PI3K/AKT/mTOR signaling.ConclusionQuercetin alleviated the progression of PMOP via activation of PI3K/AKT/mTOR signaling. Hence, this study would shed novel insights into discovering new methods against PMOP.

Myosin heavy chain-derived peptide of Gadus morhua promotes proliferation and differentiation in osteoblasts and bone formation and maintains bone homeostasis in ovariectomized mice.

Gadus morhua is an important commercial fish rich in nutrients required for daily metabolism. However, the regulation of G. morhua peptides (GMP) on osteoblast growth remains unclear. In order to clarify the regulatory effects of GMP on osteoblasts, the effects of GMP on the growth of MC3T3-E1 cells were investigated, and the osteogenic peptides were identified and screened. The results showed that GMP promoted the proliferation and differentiation of osteoblasts by regulating the BMP/WNT signaling pathway at concentrations of 1-100 μg mL-1. Molecular docking studies showed that a decapeptide, MNKKREAEFQ (P-GM-1), had a high affinity for integrins 3VI4 and 1L5G (-CDOCKER interaction energy: 161.30, 212.27 kcal mol-1). Additionally, the proliferation rate of MC3T3-E1 cells was increased by 27%, and ALP activity was significantly increased under P-GM-1 treatment (100 μg mL-1). Moreover, P-GM-1 promotes bone formation, maintains bone homeostasis, and prevents osteoporosis in ovariectomized mice by regulating the BMP/Smad signaling pathway. This study confirmed the potential of GMP in the regulation of bone mineral density and provided a certain theoretical basis for the development of anti-osteoporosis active factors from GMP.

The Effect of Apple-Derived Nanovesicles on the Osteoblastogenesis of Osteoblastic MC3T3-E1 Cells.

Osteoporosis is characterized by low bone mass and elevated structural deterioration of the bone tissue, resulting in bone weakness with an increased risk of fracture. Considering biological activities of various phytochemicals extracted from apples, we herein demonstrated the potential antiosteoporotic effects of apple-derived nanovesicles (apple NVs) using osteoblastic MC3T3-E1 cells. Apple NVs significantly stimulated the growth of MC3T3-E1 cells. The cellular alkaline phosphatase (ALP) activity was significantly upregulated in the 5 μg/mL apple NVs-treated group. In addition, the concentrarion of mineralized nodules was significantly increased in the apple NVs-treated groups. Furthermore, apple NVs increased the expression of the genes and proteins associated with osteoblast growth and differentiation, such as Runx2, ALP, OPN, and BMP2/4, which further activated ERK- and JNK-related mitogen-activated protein kinase signaling. These results demonstrate that apple NVs have a potential to prevent osteoporosis by promoting osteoblastogenesis in osteoblastic MC3T3-E1 cells through regulating the BMP2/Smad1 pathways.

Wool Keratin Nanofibers for Bioinspired and Sustainable Use in Biomedical Field.

Keratin is a biocompatible and biodegradable protein as the main component of wool and animal hair fibers. Keratin-based materials support fibroblasts and osteoblasts growth. Keratin has been extracted by sulphitolysis, a green method (no harmful chemicals) with a yield of 38-45%. Keratin has been processed into nanofibers from its solutions by electrospinning. Electrospinning is a versatile and easy-to-use technique to generate nanofibers. It is an eco-friendly and economical method for the production of randomly and uniaxially oriented polymeric nanofibers. Thanks to their high specific surface area, nanofibers have great potential in the biomedical field. Keratin nanofibers have received significant attention in biomedical applications, such as tissue engineering and cell growth scaffolds, for their biocompatibility and bio-functionality. Accordingly, we propose an extensive overview of recent studies focused on the optimization of keratinbased nanofibers, emphasizing their peculiar functions for cell interactions and the role of additive phases in blends or composite systems to particularize them as a function of specific applications (i.e., antibacterial).

Cell Type-Specific Effects of Implant Provisional Restoration Materials on the Growth and Function of Human Fibroblasts and Osteoblasts.

Implant provisional restorations should ideally be nontoxic to the contacting and adjacent tissues, create anatomical and biophysiological stability, and establish a soft tissue seal through interactions between prosthesis, soft tissue, and alveolar bone. However, there is a lack of robust, systematic, and fundamental data to inform clinical decision making. Here we systematically explored the biocompatibility of fibroblasts and osteoblasts in direct contact with, or close proximity to, provisional restoration materials. Human gingival fibroblasts and osteoblasts were cultured on the "contact" effect and around the "proximity" effect with various provisional materials: bis-acrylic, composite, self-curing acrylic, and milled acrylic, with titanium alloy as a bioinert control. The number of fibroblasts and osteoblasts surviving and attaching to and around the materials varied considerably depending on the material, with milled acrylic the most biocompatible and similar to titanium alloy, followed by self-curing acrylic and little to no attachment on or around bis-acrylic and composite materials. Milled and self-curing acrylics similarly favored subsequent cellular proliferation and physiological functions such as collagen production in fibroblasts and alkaline phosphatase activity in osteoblasts. Neither fibroblasts nor osteoblasts showed a functional phenotype when cultured with bis-acrylic or composite. By calculating a biocompatibility index for each material, we established that fibroblasts were more resistant to the cytotoxicity induced by most materials in direct contact, however, the osteoblasts were more resistant when the materials were in close proximity. In conclusion, there was a wide variation in the cytotoxicity of implant provisional restoration materials ranging from lethal and tolerant to near inert, and this cytotoxicity may be received differently between the different cell types and depending on their physical interrelationships.

Biological Effects and Molecular Mechanisms of Platelet-Rich Plasma on Periodontal Bone Regeneration

Objective: The study investigated the biological effects and molecular mechanisms of platelet-rich plasma (PRP) on periodontal bone regeneration. Methods: Electronic and manual searches were searched up to 1 October 2022 in the following databases: Pubmed, Scopus, Cochrane Library and Embase. [Platelet rich plasma or platelet or growth factors] and [periodontal] or [bone regeneration or bone defect or bone reconstruction] were used for searching. This study reviewed and analyzed published papers associated with PRP and periodontal bone defect restoration or bone regeneration or bone reconstruction. Results: Different growth factors exhibited varied biological characteristics and function. In-vitro studies, animal experiments and clinical studies confirmed that PRP displayed assorted role in periodontal bone defects repair. The growth factors secreted from PRP can promote new bone formation, soft tissue regeneration and wound healing. The fiber three-dimensional structure in PRP is conducive to the growth and migration of cells and provides strong support for the regeneration of periodontal soft and hard tissues. The anti-inflammatory characteristics of PRP are also closely related to the repair of periodontal bone defects. Conclusion: PRP played an important biological effect on periodontal bone regeneration. The mechanism is closely related to PRP promoting the growth, proliferation, differentiation and migration of periodontal ligament cells and osteoblasts, and the fiber stereo configuration of PRP and the anti-inflammatory effect of leukocytes.

Recovering Osteoblast Functionality on TiO2 Nanotube Surfaces Under Diabetic Conditions.

Titanium (Ti) and its alloys (eg, Ti6Al4V) are exceptional treatments for replacing or repairing bones and damaged surrounding tissues. Although Ti-based implants exhibit excellent osteoconductive performance under healthy conditions, the effectiveness and successful clinical achievements are negatively altered in diabetic patients. Concernedly, diabetes mellitus (DM) contributes to osteoblastic dysfunctionality, altering efficient osseointegration. This work investigates the beneficial osteogenic activity conducted by nanostructured TiO2 under detrimental microenvironment conditions, simulated by human diabetic serum. We evaluated the bone-forming functional properties of osteoblasts on synthesized TiO2 nanotubes (NTs) by anodization and Ti6Al4V non-modified alloy surfaces under detrimental diabetic conditions. To simulate the detrimental environment, MC3T3E-1 preosteoblasts were cultured under human diabetic serum (DS) of two diagnosed and metabolically controlled patients. Normal human serum (HS) was used to mimic health conditions and fetal bovine serum (FBS) as the control culture environment. We characterized the matrix mineralization under the detrimental conditions on the control alloy and the NTs. Moreover, we applied immunofluorescence of osteoblasts differentiation markers on the NTs to understand the bone-expression stimulated by the biochemical medium conditions. The diabetic conditions depressed the initial osteoblast growth ability, as evidenced by altered early cell adhesion and reduced proliferation. Nonetheless, after three days, the diabetic damage was suppressed by the NTs, enhancing the osteoblast activity. Therefore, the osteogenic markers of bone formation and the differentiation of osteoblasts were reactivated by the nanoconfigured surfaces. Far more importantly, collagen secretion and bone-matrix mineralization were stimulated and conducted to levels similar to those of the control of FBS conditions, in comparison to the control alloy, which was not able to reach similar levels of bone functionality than the NTs. Our study brings knowledge for the potential application of nanostructured biomaterials to work as an integrative platform under the detrimental metabolic status present in diabetic conditions.