Successful Regeneration Research Articles

Observations on FGF immunoreactivity in the regenerating tail blastema, and in the limb and tail scars of lizard suggest that FGFs are required for regeneration

Tail regeneration in lizards depends on the stimulation of growth factors, including Fibroblast Growth Factors (FGFs). Light and ultrastructural immunolocalization of FGFs was compared between the rege-nerating tail blastema and the limb where no regeneration occurs. A likely epithelial-mesenchymal transition occurs following amputation in both tail and limb and FGFs are present in the wound epidermis of both organs at 7-14 days post-amputation, and at lower intensity in mesenchymal cells of the blastema. Immunoreactivity for FGFs disappears in the limb wound epidermis after 14 days post-amputation and in the epithelium covering tails induced to form scars, whereas it remains in the apical tail epithelium. These observations suggest that scarring in the limb or the induced scarring in the tail correlate with the disappearance of FGFs. Basic FGF is concen-trated in the incomplete basement membrane between the epidermis and the tail blastema where the essential signaling process that allows the continuous growth of the regenerative blastema may occur. The study suggests that the successful regeneration of lizard tail is dependent on the presence of FGFs in the wound epidermis, which are probably released into the blastema.

Melatonin supplementation enhances browning suppression and improves transformation efficiency and regeneration of transgenic rough lemon plants (Citrus × jambhiri).

Enzymatic browning poses a significant challenge that limits in vitro propagation and genetic transformation of plant tissues. This research focuses on investigating how adding antioxidant substances can suppress browning, leading to improved efficiency in transforming plant tissues using Agrobacterium and subsequent plant regeneration from rough lemon (Citrus × jambhiri). When epicotyl segments of rough lemon were exposed to Agrobacterium, they displayed excessive browning and tissue decay. This was notably different from the 'Hamlin' explants, which did not exhibit the same issue. The regeneration process failed completely in rough lemon explants, and they accumulated high levels of total phenolic compounds (TPC) and polyphenol oxidase (PPO), which contribute to browning. To overcome these challenges, several antioxidant and osmoprotectant compounds, including lipoic acid, melatonin, glycine betaine, and proline were added to the tissue culture medium to reduce the oxidation of phenolic compounds and mitigate browning. Treating epicotyl segments with 100 or 200 μM melatonin led to a significant reduction in browning and phenolic compound accumulation. This resulted in enhanced shoot regeneration, increased transformation efficiency, and reduced tissue decay. Importantly, melatonin supplementation effectively lowered the levels of TPC and PPO in the cultured explants. Molecular and physiological analyses also confirmed the successful overexpression of the CcNHX1 transcription factor, which plays a key role in imparting tolerance to salinity stress. This study emphasizes the noteworthy impact of supplementing antioxidants in achieving successful genetic transformation and plant regeneration in rough lemon. These findings provide valuable insights for developing strategies to address enzymatic browning and enhance the effectiveness of plant tissue culture and genetic engineering methods with potential applications across diverse plant species.

Preventing septic implant failures in osteoporotic hip fractures using antibiotic-loaded functionalized nanocement

Osteoporosis is a prevalent skeletal disorder and the most common cause of debilitating hip fractures, primarily affecting post-menopausal women and elderly people. Implants used to stabilize osteoporotic fragility fractures are susceptible to bacterial infections, which further deteriorate the implantationsite. In the current study, we developed a nanocement-based treatment for tackling hip implant infection in an osteoporotic rat model. The development of osteoporosis was confirmed by an 80% decrease in serum estradiol levels and a further reduced T-score value of −4.170 ± 0.411 for femur neck canal. Further, an infected K-wire was implanted in femoral neck canal to induce implant-associated osteomyelitis. In order to combat infection and promote bone healing, it was followed by debridement and implantation of rifampicin-loaded nanocement functionalized with various combinations of bone marrow MSCs (bMSC) exosomes, bone morphogenetic protein-2 (BMP-2) and zoledronic acid (ZA). The findings of this study demonstrated the potential of exosomes as an alternative for reducingBMP-2 dosage, and the treatment promoted successful bone regeneration withenhanced mechanical strength, and a significant decrease in bacterial load. Improved bone remodeling and matrix deposition at the implantation site were confirmed by DXA, micro-CT, and histology. Further, immunohistochemical staining showed increased expression of collagen I and CD31, which indicated bone matrix deposition and vascularization, respectively, while reduced expression of α-SMA and NOS2 confirmed a reduction in fibrosis and inflammation, respectively. Overall, our one-step treatment strategy has the potential to prevent implant-associated bone infection in osteoporotic hip fractures, promote bone formation, and enhance the mechanical strength of osteoporotic bone, thereby preventing subsequent secondary fractures.

3D printing of gellan-dextran methacrylate IPNs in glycerol and their bioadhesion by RGD derivatives.

The ever-growing need for new tissue and organ replacement approaches paved the way for tissue engineering. Successful tissue regeneration requires an appropriate scaffold, which allows cell adhesion and provides mechanical support during tissue repair. In this light, an interpenetrating polymer network (IPN) system based on biocompatible polysaccharides, dextran (Dex) and gellan (Ge), was designed and proposed as a surface that facilitates cell adhesion in tissue engineering applications. The new matrix was developed in glycerol, an unconventional solvent, before the chemical functionalization of the polymer backbone, which provides the system with enhanced properties, such as increased stiffness and bioadhesiveness. Dex was modified introducing methacrylic groups, which are known to be sensitive to UV light. At the same time, Ge was functionalized with RGD moieties, known as promoters for cell adhesion. The printability of the systems was evaluated by exploiting the ability of glycerol to act as a co-initiator in the process, speeding up the kinetics of crosslinking. Following semi-IPNs formation, the solvent was removed by extensive solvent exchange with HEPES and CaCl2, leading to conversion into IPNs due to the ionic gelation of Ge chains. Mechanical properties were investigated and IPNs ability to promote osteoblasts adhesion was evaluated on thin-layer, 3D-printed disk films. Our results show a significant increase in adhesion on hydrogels decorated with RGD moieties, where osteoblasts adopted the spindle-shaped morphology typical of adherent mesenchymal cells. Our findings support the use of RGD-decorated Ge/Dex IPNs as new matrices able to support and facilitate cell adhesion in the perspective of bone tissue regeneration.

The molecular mechanisms underlying the regeneration process in the earthworm, Perionyx excavatus exhibit indications of apoptosis-induced compensatory proliferation (AICP).

Regeneration is a multifaceted biological phenomenon that necessitates the intricate orchestration of apoptosis, stem cells, and immune responses, culminating in the regulation of apoptosis-induced compensatory proliferation (AICP). The AICP context of research is observed in many animal models like in Hydra, Xenopus, newt, Drosophila, and mouse but so far not reported in earthworm. The earthworm Perionyx excavatus is used in the present study to understand the relationship between AICP-related protein expression and regeneration success in different conditions (normal regeneration and abnormal multiple bud formation). Initially, the worms are amputated into five equal portions and it is revealed that regeneration in P. excavatus is clitellum independent and it gives more preference for anterior regeneration (regrowth of head portion) than for posterior regeneration (regrowth of tail portion). The posterior segments of the worm possess enormous regeneration ability but this is lacking in anterior segments. Alkaline phosphate, a stem cell marker, shows strong signals throughout all the posterior segments but it decreases in the initial 1st to 15th anterior segments which lack the regeneration ability. While regenerating normally, it was suggested that the worm follow AICP principles. This is because there was increased expression of apoptosis signals throughout the regeneration process along with constant expression of stem cell proliferation response together with cellular proliferation. In amputated posterior segments maintained in vitro, the apoptosis signals were extensively detected on the 1st day. However, on the 4th and 6th days, caspase-3 and H2AX expression are significantly suppressed, which may eventually alter the Wnt3a and histone H3 patterns that impair the AICP and result in multiple bud formation. Our results suggest that AICP-related protein expression pattern is crucial for initiating proper regeneration.

3D-printed PCL framework assembling ECM-inspired multi-layer mineralized GO-Col-HAp microscaffold for in situ mandibular bone regeneration

BackgroundIn recent years, natural bone extracellular matrix (ECM)-inspired materials have found widespread application as scaffolds for bone tissue engineering. However, the challenge of creating scaffolds that mimic natural bone ECM’s mechanical strength and hierarchical nano-micro-macro structures remains. The purposes of this study were to introduce an innovative bone ECM-inspired scaffold that integrates a 3D-printed framework with hydroxyapatite (HAp) mineralized graphene oxide-collagen (GO-Col) microscaffolds and find its application in the repair of mandibular bone defects.MethodsInitially, a 3D-printed polycaprolactone (PCL) scaffold was designed with cubic disks and square pores to mimic the macrostructure of bone ECM. Subsequently, we developed multi-layer mineralized GO-Col-HAp microscaffolds (MLM GCH) to simulate natural bone ECM's nano- and microstructural features. Systematic in vitro and in vivo experiments were introduced to evaluate the ECM-inspired structure of the scaffold and to explore its effect on cell proliferation and its ability to repair rat bone defects.ResultsThe resultant MLM GCH/PCL composite scaffolds exhibited robust mechanical strength and ample assembly space. Moreover, the ECM-inspired MLM GCH microscaffolds displayed favorable attributes such as water absorption and retention and demonstrated promising cell adsorption, proliferation, and osteogenic differentiation in vitro. The MLM GCH/PCL composite scaffolds exhibited successful bone regeneration within mandibular bone defects in vivo.ConclusionsThis study presents a well-conceived strategy for fabricating ECM-inspired scaffolds by integrating 3D-printed PCL frameworks with multilayer mineralized porous microscaffolds, enhancing cell proliferation, osteogenic differentiation, and bone regeneration. This construction approach holds the potential for extension to various other biomaterial types.Graphical

CD38 promotes hematopoietic stem cell dormancy.

A subpopulation of deeply quiescent, so-called dormant hematopoietic stem cells (dHSCs) resides at the top of the hematopoietic hierarchy and serves as a reserve pool for HSCs. The state of dormancy protects the HSC pool from exhaustion throughout life; however, excessive dormancy may prevent an efficient response to hematological stresses. Despite the significance of dHSCs, the mechanisms maintaining their dormancy remain elusive. Here, we identify CD38 as a novel and broadly applicable surface marker for the enrichment of murine dHSCs. We demonstrate that cyclic adenosine diphosphate ribose (cADPR), the product of CD38 cyclase activity, regulates the expression of the transcription factor c-Fos by increasing the release of Ca2+ from the endoplasmic reticulum (ER). Subsequently, we uncover that c-Fos induces the expression of the cell cycle inhibitor p57Kip2 to drive HSC dormancy. Moreover, we found that CD38 ecto-enzymatic activity at the neighboring CD38-positive cells can promote human HSC quiescence. Together, CD38/cADPR/Ca2+/c-Fos/p57Kip2 axis maintains HSC dormancy. Pharmacological manipulations of this pathway can provide new strategies to improve the success of stem cell transplantation and blood regeneration after injury or disease.

The Source of Melanocytes in Ortho- and Heterotopic Tail Regenerates of Axolotls and the Dependence of the Regenerative Response on the Presence of Neural Tissue

We studied the regeneration of orthotopic and heterotopic tails in larval axolotls. First, we analyzed tail regeneration following reciprocal exchange of cuffs of tail integument between dark-colored (wild-type) and yellow-colored (hybrid) larval animals. Second, we studied tail regeneration in larval axolotls following transplantation of cuffs of tail integument from metamorphosed dark-colored conspecifics and from an adult fire salamander. In all cases, the amputation planes involved the transplanted integumental cuffs. In the first experiment, the regenerated tails showed the color of the host animals, not that of the transplanted cuffs. This suggests that the melanocytes of the regenerated tails were derived from the host hypodermis. Following transplantation of metamorphosed skin from axolotls and a fire salamander onto larval axolotls, the metamorphosed epidermis reverted to a larval condition. This indicates that the state of differentiation of the metamorphosed epidermis was not permanent. Rather, in order to maintain the metamorphosed epidermal structure, a continuous exposure of the animals to sufficient levels of thyroid hormones was required. Transplantation of tail buds from yellow-colored onto dark-colored axolotl embryos caused the formation of yellow-colored tails both in the head and the anterior limb region of the hosts. Incomplete resection of these heterotopic tails was followed by tail regeneration, while no tail regeneration occurred following complete resection of the heterotopic tails. Successful tail regeneration depended on the presence of neural tissue along the resection plane.

Pro-vasculogenic Fibers by PDA-Mediated Surface Functionalization Using Cell-Free Fat Extract (CEFFE).

Formation of adequate vascular network within engineered three-dimensional (3D) tissue substitutes postimplantation remains a major challenge for the success of biomaterials-based tissue regeneration. To better mimic the in vivo angiogenic and vasculogenic processes, nowadays increasing attention is given to the strategy of functionalizing biomaterial scaffolds with multiple bioactive agents. Aimed at engineering electrospun biomimicking fibers with pro-vasculogenic capability, this study was proposed to functionalize electrospun fibers of polycaprolactone/gelatin (PCL/GT) by cell-free fat extract (CEFFE or FE), a newly emerging natural "cocktail" of cytokines and growth factors extracted from human adipose tissue. This was achieved by having the electrospun PCL/GT fiber surface coated with polydopamine (PDA) followed by PDA-mediated immobilization of FE to generate the pro-vasculogenic fibers of FE-PDA@PCL/GT. It was found that the PDA-coated fibrous mat of PCL/GT exhibited a high FE-loading efficiency (∼90%) and enabled the FE to be released in a highly sustained manner. The engineered FE-PDA@PCL/GT fibers possess improved cytocompatibility, as evidenced by the enhanced cellular proliferation, migration, and RNA and protein expressions (e.g., CD31, vWF, VE-cadherin) in the human umbilical vein endothelial cells (huvECs) used. Most importantly, the FE-PDA@PCL/GT fibrous scaffolds were found to enormously stimulate tube formation in vitro, microvascular development in the in ovo chick chorioallantoic membrane (CAM) assay, and vascularization of 3D construct in a rat subcutaneous embedding model. This study highlights the potential of currently engineered pro-vasculogenic fibers as a versatile platform for engineering vascularized biomaterial constructs for functional tissue regeneration.

Bioactive Glass and Silica Particles for Skeletal and Cardiac Muscle Tissue Regeneration.

When skeletal and cardiac tissues are damaged, surgical approaches are not always successful and tissue regeneration approaches are investigated. Reports in the literature indicate that silica nanoparticles and bioactive glasses (BGs), including silicate bioactive glasses (e.g., 45S5 BG), phosphate glass fibers, boron-doped mesoporous BGs, borosilicate glasses, and aluminoborates, are promising for repairing skeletal muscle tissue. Silica nanoparticles and BGs have been combined with polymers to obtain aligned nanofibers and to maintain controlled delivery of nanoparticles for skeletal muscle repair. The literature indicates that cardiac muscle regeneration can be also triggered by the ionic products of BGs. This was observed to be due to the release of vascular endothelial growth factor and other growth factors from cardiomyocytes, which regulate endothelial cells to form capillary structures (angiogenesis). Specific studies, including both in vitro and in vivo approaches, are reviewed in this article. The analysis of the literature indicates that although the research field is still very limited, BGs are showing great promise for muscle tissue engineering and further research in the field should be carried out to expand our basic knowledge on the application of BGs in muscle (skeletal and cardiac) tissue regeneration. Impact statement This review highlights the potential of silica particles and bioactive glasses (BGs) for skeletal and cardiac tissue regeneration. These biomaterials create scaffolds triggering muscle cell differentiation. Ionic products from BGs stimulate growth factors, supporting angiogenesis in cardiac tissue repair. Further research is required to expand our know-how on silica particles and BGs in muscle tissue engineering.

Assessing the functional potential of conditioned media derived from amniotic epithelial stem cells engineered on 3D biomimetic scaffolds: An in vitro model for tendon regeneration

Tendon diseases pose a significant challenge in regenerative medicine due to the limited healing capacity of this tissue. Successful tendon regeneration requires a combination of angiogenesis, immune response, and tenogenesis processes. An effective tendon engineering (TE) strategy must finely tune this systems’ interplay toward homeostasis.This study explores in vitro the paracrine influence of amniotic epithelial stem cells (AECs) engineered on a validated 3D electrospun PLGA scaffolds on HUVECs (angiogenesis), PBMCs/Jurkat (immune response), and AECs (tenogenic stem cell activation).The results revealed the role of scaffold's topology and topography in significantly modulating the paracrine profile of the cells. In detail, AECs basal release of bioactive molecules was boosted in the cells engineered on 3D scaffolds, in particular VEGF-D, b-FGF, RANTES, and PDGF-BB (p < 0.0001 vs. CMCTR). Moreover, biological tests demonstrated 3D scaffolds' proactive role in potentiating AECs' paracrine inhibition on PBMCs proliferation (CM3Dvs. CTR, p < 0.001) and LPS-mediated Jurkat activation with respect to controls (CM3D and CM2Dvs. CTR, p < 0.01 and p < 0.05, respectively), without exerting any in vitro pro-angiogenic role in promoting HUVECs proliferation and tubule formation. Teno-inductive paracrine ability of AECs engineered on 3D scaffolds was assessed on co-cultured ones, which formed tendon-like structures. These latter demonstrated an upregulation of tendon-related genes (SCX, THBS4, COL1, and TNMD) and the expression TNMD and COL1 proteins.Overall, this research underscores the pivotal role of the 3D topology and topography of PLGA tendon mimetic scaffolds in orchestrating effective tendon regeneration through modulating cell behavior and crosstalk between engineered stem cells and different subpopulations in the damaged tendon.

Guava seed activated carbon loaded calcium alginate aerogel for the adsorption of diclofenac sodium: Characterization, isotherm, kinetics, and optimization via Box-Behnken design

This study aimed to develop a novel adsorbent designed for the removal of diclofenac sodium (DS) from water. The synthesized adsorbent, a composite sponge known as guava seeds activated carbon loaded calcium alginate (GSAC@CA aerogel), was created through the combination of powdered activated carbon derived from guava seeds and loaded onto a calcium alginate hydrogel. Characterization through SEM, XRD, FT-IR, BET, and XPS revealed a confirmed surface area of 738.82 m2/g. The investigation delved into assessing the influence of pH, initial DS concentration, and adsorbent dose on the adsorption of DS. Isotherm studies on adsorption suggested that a Langmuir model provided a good fit, indicating a monolayer adsorption process. Kinetic studies revealed a well-fitted pseudo-second-order model, shedding light on the dynamics of the reaction. The chemisorption nature was elucidated by the Dubinin-Radushkevich model, demonstrating an adsorption energy of 22.6 kJ/mol. These results affirm the potential of the GSAC@CA aerogel composite sponge as an efficient adsorbent for removing diclofenac sodium from water. Examination of the impact of temperature on the adsorption process revealed an endothermic behavior, indicating an increase in temperature. The positive change in entropy suggested the spontaneous nature of the reaction. Remarkably, the GSAC@CA aerogel composite sponge exhibited strong adsorption capabilities, achieving a maximum adsorption capacity of 489.97 mg/g. Across five consecutive cycles, the composite consistently demonstrated high-level adsorption, maintaining a removal efficiency of 87.77 %. The adsorption mechanism of diclofenac sodium (DS) on the GSAC@CA aerogel composite sponge was determined to encompass various processes, such as hydrogen bonding, π-π interactions, ion exchange, and electrostatic pore filling. Additionally, the adsorbent demonstrated successful regeneration over three cycles when applied to a real-world sample. The incorporation of the Box-Behnken design (BBD) introduced a strategic aspect to enhance adsorption outcomes, offering valuable insights for optimizing the adsorption process in practical applications.

Enhancing the regeneration of TIS-derived somatic embryos of oil palm (Elaeis guineensis Jacq.) with CaCl2 and GA3

Micropropagation of oil palm through somatic embryogenesis (SE) technique provides clonal and uniform seedlings as high-quality planting materials for plantation. However, a problem associated with oil palm SE is the low regeneration rate of the somatic embryos. It was revealed that nutrient balance played a vital role in the success of SE regeneration. Therefore, a study was conducted to determine the balance between a macronutrient, calcium, and the germination-inducing hormone, Gibberellin (GA3) in enhancing the regeneration of the oil palm somatic embryos derived from the Temporary Immersion System (TIS). Treatments consist of various concentrations of CaCl2 (440, 880, or 1320 mg L-1) and enrichment of GA3 (2 or 4 mg L-1) applied into SE regeneration media, followed by the observation of regeneration rate and shoot tip necrosis after five weeks of culture. The study revealed that CaCl2 in the concentration of 880 mg L-1 was the best in stimulating oil palm TIS-derived SE regeneration. A higher concentration of GA3 (4 mg L-1) was needed in the first transition of SE from TIS into SE regeneration media in solid culture, while further SE-derived shoots development needed a lower concentration of GA3.

Reduced and modified graphene oxide with Ag/V2O5 as a ternary composite visible light photocatalyst against dyes and pesticides

Water pollution by dyes and pesticides poses significant threats to our ecosystem. In this research, a visible-light ternary composite photocatalytic system was fabricated using graphene oxide (GO) by reducing with N2H4, modifying with KOH, and decorating with Ag/V2O5. The fabricated photocatalysts were characterized through FTIR, SEM, XRD, BET, PL, EDX, ESR, UV–vis spectroscopy, TGA, ESI-MS, and Raman spectroscopy. The point zero charge of the reduced and modified GO (RMGO/Ag/V2O5) was measured to be 6.7 by the pH drift method. This ternary composite was able to achieve complete removal of methyl orange (MO) and chlorpyrifos (CP) in solutions in 80 min under the optimum operation conditions (e.g., in terms of pollutant/catalyst concentrations, pH effects, and contact time). The role of active species responsible for photocatalytic activity was confirmed by scavenger analysis and ESR investigations. The potential mechanism for photocatalytic activity was studied through a fragmentation process carried out by MS analysis. Through nonlinear fitting of the experimental data, MO and CP exhibited the best fit results with the pseudo 1st-order kinetics (quantum yields of 1.07 × 10−3 and 2.16 × 10−3 molecules photon−1 and space-time yields of 1.53 × 10−5 and 2.7 × 10−5 molecules photon−1 mg−1, respectively). The structure of the nanomaterials remained mostly intact to support increased stability and reusability of the prepared photocatalysts even after 10 successive regeneration cycles.

In Vitro Studies on 3D-Printed PLA/HA/GNP Structures for Bone Tissue Regeneration.

The successful regeneration of large-size bone defects remains one of the most critical challenges faced in orthopaedics. Recently, 3D printing technology has been widely used to fabricate reliable, reproducible and economically affordable scaffolds with specifically designed shapes and porosity, capable of providing sufficient biomimetic cues for a desired cellular behaviour. Natural or synthetic polymers reinforced with active bioceramics and/or graphene derivatives have demonstrated adequate mechanical properties and a proper cellular response, attracting the attention of researchers in the bone regeneration field. In the present work, 3D-printed graphene nanoplatelet (GNP)-reinforced polylactic acid (PLA)/hydroxyapatite (HA) composite scaffolds were fabricated using the fused deposition modelling (FDM) technique. The in vitro response of the MC3T3-E1 pre-osteoblasts and RAW 264.7 macrophages revealed that these newly designed scaffolds exhibited various survival rates and a sustained proliferation. Moreover, as expected, the addition of HA into the PLA matrix contributed to mimicking a bone extracellular matrix, leading to positive effects on the pre-osteoblast osteogenic differentiation. In addition, a limited inflammatory response was also observed. Overall, the results suggest the great potential of the newly developed 3D-printed composite materials as suitable candidates for bone tissue engineering applications.

Periapical bacterial disinfection is critical for dental pulp regenerative cell therapy in apical periodontitis in dogs

BackgroundApplication of pulp regenerative cell therapy for mature teeth with periapical lesions is a critical clinical challenge. The bacterial infection in inaccessible location within the root canal system and in the periapical lesions could cause resistance and impediment, leading to limitations in successful therapy. Thus, the aim of this study was to examine the effect of residual bacteria on the outcome of pulp regeneration in mature teeth with apical periodontitis in dogs.MethodsPeriapical lesions were induced in 32 root canals of 4 dogs in two different models in severities, model A and model B. Model A (moderate infection): the canal exposed to the oral cavity for 2 weeks and then closed for 2 weeks. Model B (severe infection): the canal exposed to the oral cavity for 2 months and then closed for 5 months. All root canals were irrigated with 6% sodium hypochlorite, and 3% EDTA and further with 0.015% levofloxacin-containing nanobubbles, which was also used as an intracanal medicament. The aseptic conditions were examined by bacterial anaerobic culture and/or PCR analyses. The root canal treatment was repeated several times, and allogeneic dental pulp stem cells were transplanted into the root canals. The radiographic evaluation of periapical lesions was performed by cone-beam computed tomography before the first treatment, just after cell transplantation, and after 2 months and 6 months in both model A, model B, respectively. The animals were then sacrificed and the jaw blocks were harvested for histological and histobacteriological evaluations of pulp regeneration and periapical tissue healing. Furthermore, the DiI-labelled DPSCs were transplanted into the root canals after complete disinfection (n = 4) or without root canal treatment (n = 4) in the apical periodontitis model (model A) in one dog, and cell localization was compared 72 h after transplantation.ResultsIn 8 out of 12 canals from model A, and 10 out of 15 canals from model B, pulp regeneration with good vascularization, innervation, and a significant reduction in the radiolucent area of the periapical lesions were observed. However, in the other 4 canals and 5 canals from model A and model B, respectively, no pulp tissue was regenerated, and inflammation in the periapical tissue, and external resorption or healed external resorption were detected. The presence of residual bacteria in the periapical tissues and severe inflammation were significantly associated with inhibition of regenerated pulp tissue in these 9 unsuccessful canals (P < 0.05, each) (OR = 0.075, each) analyzed by multiple logistic regression analysis. For cellular kinetics, transplanted cells remained in the disinfected root canals, while they were not detected in the infected root canals, suggesting their migration through the apical foramen under the influence of inflammation.ConclusionsA true pulp-dentin complex was regenerated in the root canal by the pulp regenerative therapy in mature teeth with apical lesions. The successful pulp regeneration was negatively associated both with residual bacteria and inflammation in the periapical tissue.

Allogeneic Bone Marrow Mesenchymal Stromal Cell Transplantation Induces Dentin Pulp Complex-like Formation in Immature Teeth with Pulp Necrosis and Apical Periodontitis

IntroductionDental pulp regeneration is challenging in endodontics. Cellular therapy is an alternative approach to induce dental pulp regeneration. Mesenchymal stromal cells (MSCs) have the capacity to induce dental pulp-like tissue formation. In this study, we evaluated the capacity of allogeneic bone marrow MSCs (BM-MSCs) to regenerate pulp following necrosis and apical periodontitis in children's permanent immature apex teeth. MethodsPatients aged 8 to 12 years with pulp necrosis and apical periodontitis were evaluated. The study included 15 teeth (13 incisors and 2 molars) from 14 patients (8 boys and 6 girls). Radiographic evaluation showed periapical radiolucency and immature apex teeth. There was no response to cold or electric pulp testing. The root canal of each tooth was cleaned, shaped, and Ca(OH)2 used as an interappointment medication. Cryopreserved allogeneic BM-MSCs were thawed, expanded, incorporated into preclotted platelet-rich plasma, and implanted into the tooth's pulp cavity. They were sealed with bioceramic cement and composite. Sensibility, apical foramen, calcium deposits within the root canal, and resolution of periapical lesions were evaluated in each tooth over the following 12 months. ResultsBased on 9 variables established for dental pulp-like tissue regeneration, all MSC-treated teeth showed evidence of successful regeneration. Clinical and radiographic evaluation of the treated teeth showed periapical lesion healing, sensitivity to cold and electricity, decreased width of the apical foramen, and mineralization within the canal space. ConclusionsTransplantation of allogeneic MSCs induces the formation of dental pulp-like tissue in permanent immature apex teeth with pulp necrosis and apical periodontitis. Implant of MSCs constitutes a potential therapy in regenerative endodontics in pediatric dentistry. Future studies incorporating a larger sample size may confirm these results.

The Last Molar - Entire Pad Preservation Technique (L-EPPT) as a Regenerative Surgical Approach for Combined Distal Intrabony Defects and Furcation Involvement: Case Reports.

Primary wound closure is the most important factor in successful periodontal tissue regeneration when using biomaterials. However, in the distal region of the last molar, there is concern that direct surgical access to an intraosseous defect may induce the leakage of biomaterials, impair the blood supply, and make healing difficult. This case series introduces the last molar-entire pad preservation technique (L-EPPT), which was designed to preserve the gingiva of the last molar centrum and secure the operative field of the furcation and distal bone defects, thereby providing an optimal environment for wound healing for regenerative therapy. This technique for preservation of the distal gingiva was applied in two cases involving the combination of a buccal class II furcation defect and a 2-3-wall intrabony defect in the last molar for the use of a combination of bone graft and enamel matrix derivatives in periodontal regeneration. Clinical outcomes were recorded at 18 months (case 1) and 8 months (case 2) after surgery. In both cases, the class II furcation defects were completely closed, and the probing pocket depth was improved to < 3 mm with no bleeding on probing. Further research is warranted to verify the efficacy of this technique.

Integrated Ozonation Ni-NiO/Carbon/g-C3N4 Nanocomposite-Mediated Catalytic Decomposition of Organic Contaminants in Wastewater under Visible Light.

Developing a hybrid process for wastewater purification is of utmost importance to make conventional methods more efficient and faster. Herein, an effective visible light-active nickel-nickel oxide/carbon/graphitic carbon nitride (Ni-NiO/C/g-C3N4)-based nanocatalyst was developed. A hybrid process based on ozonation and Ni-NiO/C/g-C3N4 visible light photocatalysis was applied to decolourize the Congo red (CR), Alizarin Red S (ARS), and real dairy industry wastewater. The synthesized catalyst was characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), Χ-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and UV-Vis diffuse reflectance spectrophotometry (UV-Vis DRS). The factors affecting the catalytic process were evaluated, including contact time, solution pH, initial dye concentration, etc. The degradation rate of CR and ARS was compared between the photocatalysis, ozonation, and integrated photocatalytic ozonation (PC/O3) methods. The results showed 100% degradation of CR and ARS within 5 min and 40 min, respectively, by integrated PC/O3. The reusability of the modified catalyst was evaluated, and four successive regenerations were achieved. The modified Ni-NiO/C/g-C3N4 composite could be considered an effective, fast, and reusable catalyst in an integrated PC/O3 process for the complete decolourization of wastewater.

Achieving the ideal balance between biological and mechanical requirements in composite bone scaffolds through a voxel-based approach

Achieving successful bone regeneration necessitates the design of scaffolds that meet diverse biological and mechanical requirements, often leading to conflicts in the design parameters. A key conflict arises between scaffold porosity and stiffness. Increasing porosity facilitates cell infiltration and nutrient exchange, promoting bone regeneration. However, higher porosity compromises scaffold stiffness, which is crucial for providing structural support in the defective region. Furthermore, appropriate scaffold stiffness is crucial for preventing stress shielding. Conventional geometry-based design methods utilizing single-phase materials have limited flexibility in resolving such conflicts. To address this challenge, we propose a voxel-based method for designing composite scaffolds composed of hydroxyapatite (HA) and polylactic acid (PLA). Our strategy involves first satisfying primary biological requirements by selecting appropriate porosity, pore shape, and size. Subsequently, scaffold stiffness requirements are met by selecting suitable phase materials and tuning their contents. The study demonstrates that the voxel-based approach effectively balances both biological and mechanical requirements in scaffold design. This method addresses the limitations of traditional designs by achieving an optimal balance between porosity and stiffness, which is crucial for scaffold performance in biomedical applications. Moreover, the scaffolds designed using this method can be manufactured using voxel-based 3D printing technology, which is emerging in the field. Future advancements in voxel-based 3D printing technology will further enhance the feasibility and practicality of this approach for bone tissue engineering applications.