Effects of exosomes derived from platelet-rich plasma on osteogenic differentiation of dental pulp stem cells.

Platelet-rich plasma (PRP) can cause osteogenic differentiation of dental pulp stem cells (DPSCs). However, the effect of exosomes derived from PRP (PRP-Exos) on osteogenic differentiation of DPSCs remains unclear. Herein, we evaluated the impact of PRP-Exos on osteogenic differentiation of DPSCs. PRP-Exos were isolated and identified by transmission electron microscopy (TEM) and western blotting (WB). Immunofluorescence staining was performed to evaluate endocytosis of PRP-Exos by DPSCs. Alkaline phosphatase staining, alizarin red staining, western blot and qRT-PCR were carried out to evaluate the DPSCs osteogenic differentiation. The sequencing microRNA (miRNA) was conducted to determine the microRNA profile of PRP-Exos treated and untreated DPSCs. The results showed that endocytosis of PRP-Exos stimulated DPSCs odontogenic differentiation by elevated expression of ALP, DMP-1, OCN, and RUNX2. ALP activity and calcified nodules formation of PRP-Exos treated DPSCs were considerably elevated relative to that of the control group. MicroRNA sequencing revealed that 112 microRNAs considerably varied in PRP-Exos treated DPSCs, of which 84 were elevated and 28 were reduced. Pathway analysis suggested that genes targeted by differentially expressed (DE) miRNAs were contributed to many signaling cascades, such as the Wnt cascade. 65 genes targeted by 30 DE miRNA were contributed to Wnt signaling. Thus, it can be infered that PRP-Exos could enhance osteogenic differentiation and alter the miRNA expression profile of DPSCs.

Oral microbial changes and oral disease management before and after the treatment of hematological malignancies: a narrative review.

OBJECTIVES: Patients with hematological malignancies have dynamic changes in oral microbial communities before and after treatment. This narrative review describes the changes in oral microbial composition and diversity, and discusses an oral microbe-oriented strategy for oral disease management. MATERIALS AND METHODS: A literature search was performed in PubMed/Medline, Web of Science, and Embase for articles published between 1980 and 2022. Any articles on the changes in oral microbial communities in patients with hematological malignancies and their effects on disease progression and prognosis were included. RESULTS: Oral sample detection and oral microbial sequencing analysis of patients with hematological malignancies showed a correlation between changes in oral microbial composition and diversity and disease progression and prognosis. The possible pathogenic mechanism of oral microbial disorders is the impairment of mucosal barrier function and microbial translocation. Probiotic strategies, antibiotic strategies, and professional oral care strategies targeting the oral microbiota can effectively reduce the risk of oral complications and the grade of severity in patients with hematological malignancies. CLINICAL RELEVANCE: This review provides dentists and hematologists with a comprehensive understanding of the host-microbe associated with hematologic malignancies and oral disease management advice.

The prevalence of gingivitis and related risk factors in schoolchildren aged 6-12 years old.

BACKGROUND: According to epidemiological studies, gingivitis is a common disease. However, its morbidity, considerably varies among individual. This study aimed to investigate the epidemiological characteristics of gingivitis, including prevalence, severity, intraoral distribution, and associated risk factors, in children aged 6-12 years in Jinzhou, China. METHODS: A multistage, whole-group, randomized sample of 2880 children aged 6-12 years in Jinzhou City, China, was selected and clinically examined. Each selected child completed a questionnaire on sociodemographic factors and oral health behaviors in cooperation with the investigator and teacher. Gingival bleeding refers to the bleeding of 10% or more teeth under the condition of ingivitis. Gingivitis was further categorized into localized gingivitis (30% ≥ number of teeth positive for gingival bleeding ≥ 10%) and generalized gingivitis (number of positive for gingival bleeding > 30%). The score of gingival bleeding was recorded using the Gingival Index. RESULTS: The prevalence of gingivitis in children aged 6-12 years in Jinzhou was 28.58%, including 701 cases of localized gingivitis (24.3%) and 122 cases of generalized gingivitis (4.2%). There were 429 cases (28.3%) of gingivitis in males and 394 cases (28.9%) in females, with no statistically significant difference in prevalence between males and females (P > 0.05). Chi-square tests and binary logistic regression analysis showed that aging, dental calculus, plaque, and dental crowding were significantly associated with a high prevalence of gingivitis. CONCLUSIONS: Our study showed that dental calculus, large amount dental plaque, poor oral health behavior, and oral health awareness are associated with the prevalence of gingivitis and maintaining children's oral health requires professional guidance and regular preventive care.

Effect of Comonomers on Physical Properties and Cell Attachment to Silica-Methacrylate/Acrylate Hybrids for Bone Substitution.

Hybrids with a silica network covalently bonded to a polymer are promising materials for bone repair. Previous work on synthesizing methyl methacrylate (MMA) based copolymers by reversible addition-fragmentation chain transfer (RAFT) polymerization gives high tailorability of mechanical properties since sophisticated polymer structures can be designed. However, more flexible hybrids would be beneficial. Here, n-butyl methacrylate (BMA) and methyl acrylate (MA) based hybrids are produced. Unlike MMA, BMA and MA hybrids do not show plastic deformation, and BMA hybrid has strain to failure of 33%. Although the new hybrids are more flexible, preosteoblast cells do not adhere on their surfaces, due to higher hydrophobicity and lower stiffness. Comonomer choice is crucial for bone regenerative hybrids.

The Prevalence of Dental Carious Lesions and Associated Risk Factors in Chinese Children Aged 7-9 Years.

PURPOSE: To investigate the prevalence, severity, oral distribution, and associated risk factors of carious lesions in the pri- mary teeth in children in Jinzhou, China, aged 7-9 years. MATERIALS AND METHODS: A total of 1603 primary school students aged 7-9 years old from public and private schools in Jinzhou were recruited using multi-stage, stratified, and random sampling methods for cross-sectional studies. Carious lesions in the primary teeth of school-age children were detected and recorded according to the World Health Organiza- tion standard, and a questionnaire was collected from a parent or guardian with information on the relevant risk factors for the child. Odds ratios and 95% confidence intervals of factors related to carious lesions were estimated using binary logistic regression analysis (p<0.05). RESULTS: The prevalence of carious lesions in the primary teeth was 74.5%, the average number of carious lesions was 3.02, and dmft was 4.08 ± 2.74. There were 655 cases (77.1%) of dental carious lesions in boys and 546 cases (72.5%) in girls, and the difference between them was statistically significant (p<0.05). Binary logistic regression analysis showed that the mother's educational level, brushing frequency, brushing time, and consumption of soft drinks, desserts, and sweets were all associated with a higher prevalence of carious lesions (p<0.05). CONCLUSIONS: The children in our sample had a high incidence of carious lesions of the primary teeth, especially the man- dibular primary molars. Social demographic factors, oral hygiene habits, and dietary habits all play an important role in the occurrence of carious lesions.

High concentrations of NaF aggravate periodontitis by promoting M1 polarization in macrophages.

High-concentration fluoride treatment is commonly used to prevent dental caries in the oral cavity, and fluorine-containing protective paint is used to alleviate common root sensitivity symptoms in patients with periodontitis after periodontal treatment. Recent studies have confirmed its safe use in normal oral environments. However, whether fluoride treatment affects the progression of periodontitis in an inflammatory microenvironment remains unclear. Immunometabolism is crucial for maintaining bone regeneration and repair in periodontitis, and the precise regulation of macrophage polarisation is crucial to this process. Fluoride can influence the immune microenvironment of bone tissue by regulating immune metabolic processes. Herein, we investigated the effects of high concentrations of sodium fluoride (NaF) on periodontal tissues. We examined the expression of osteogenic and M1/M2 macrophage polarisation markers and glucose metabolism in macrophages. RNA sequencing was used to study differentially expressed genes related to M1 polarisation and glucose metabolism in treated macrophages. The results showed that NaF indirectly affects human periodontal ligament cells (hPDLCs), aggravating bone loss, tissue destruction, and submandibular lymph node drainage. Furthermore, NaF promoted glycolysis in macrophages and M1 polarisation while inhibiting osteogenic differentiation. These findings suggest that NaF has a direct effect on hPDLCs. Moreover, we found that high concentrations of NaF stimulated M1 polarisation in macrophages by promoting glycolysis. Overall, these results suggest that M1 macrophages promote the osteoclastic ability of hPDLCs and inhibit their osteogenic ability, eventually aggravating periodontitis. These findings provide important insights into the mechanism of action of NaF in periodontal tissue regeneration and reconstruction, which is critical for providing appropriate recommendations for the use of fluoride in patients with periodontitis.

Prevalence and Influencing Factors of Mixed Dentition Malocclusion in Children Aged 6-12 Years in Jinzhou, China.

PURPOSE: To investigate the prevalence, clinical manifestations and related risk factors of malocclusion in schoolchildren of Jinzhou City, China. MATERIALS AND METHODS: A total of 2162 children aged 6-12 years were randomly selected from various districts of Jinzhou. Conventional clinical examination was performed by stomatologists, and the results were described based on different clinical manifestations of malocclusion and individual normal occlusion. Further, a questionnaire survey completed by children's parents or guardians provided the demographic data, lifestyle, and oral habits. The distribution of individual normal occlusion and malocclusion was documented in percentage, and Pearson's Χ2 was used for two-factor analysis. The data were statistically analysed using SPSS software (version 25.0) with a significance level of α = 0.05. RESULTS: A total of 1129 boys and 1033 girls were included in this study, i.e. 52.2% and 47.8% of the total number of children, respectively. The prevalence of malocclusion in children aged 6-12 years old in Jinzhou was 67.9%, of which crowded dentition was the most common form, with a prevalence of 71.8%, followed by deep overbite, anterior crossbite, dental spacing, deep overjet, anterior edge-to-edge occlusion, and anterior open bite. In the logistic regression model, the results showed that BMI index had little effect on the occurrence of malocclusion (p > 0.05), while dental caries, bad oral habits, retained primary teeth, and a low labial frenum were all related to the occurrence of malocclusion (p < 0.05). Moreover, the higher frequency and duration of bad oral habits were associated with a higher likelihood of malocclusion. CONCLUSIONS: The prevalence of malocclusion in children aged 6-12 years in Jinzhou is high. In addition, bad oral habits (such as lip biting, tongue thrusting, biting/gnawing objects, unilateral chin supporting, and unilateral mastication) and other related risk factors (such as dental caries, mouth breathing, retention of primary teeth, and low labial frenum, etc) were associated with malocclusion.

Hsa_Circ_0005044 Promotes Osteo/Odontogenic Differentiation of Dental Pulp Stem Cell Via Modulating miR-296-3p/FOSL1.

Circular RNAs (circRNAs) are a form of RNAs that lack coding potential. The role of such circRNAs in dental pulp stem cell (DPSC) osteo/odontogenic differentiation remains to be determined. In this study, circRNA expression profiles in DPSC osteo/odontogenic differentiation process were analyzed by RNA-seq. qRT-PCR was used to confirm the differential expression of circ_0005044, miR-296-3p, and FOSL1 in DPSC osteogenic differentiation process. Circ_0005044, miR-296-3p, and FOSL1 were knocked down or overexpressed. Osteoblastic activity and associated mineral activity were monitored via alkaline phosphatase (ALP) and alizarin red S (ARS) staining. Interactions between miR-296-3p, circ_0005044, and FOSL1 were assessed through luciferase reporter assays. Finally, an in vivo system was used to confirm the relevance of circ_0005044 to osteoblastic differentiation. As results, we detected significant circ_0005044 and FOSL1 upregulation in DPSC osteo/odontogenic differentiation process, as well as concomitant miR-296-3p downregulation. When knocking down circ_0005044 or overexpressed miR-296-3p, this significantly inhibited osteogenesis. Luciferase reporter assay confirmed that miR-296-3p was capable of binding to conserved sequences in the wild-type forms of both the circ_0005044 and FOSL1. Furthermore, knocking down circ_0005044 in vivo significantly attenuated bone formation. Therefore, the circ_0005044/miR-2964-3p/FOSL1 axis regulates DPSC osteo/odontogenic differentiation, which may provide potential molecular targets for dental-pulp complex regeneration.

A Conductive Bioengineered Cardiac Patch for Myocardial Infarction Treatment by Improving Tissue Electrical Integrity.

Conductive scaffolds are of great value for constructing functional myocardial tissues and promoting tissue reconstruction in the treatment of myocardial infarction (MI). Here, a novel scaffold composed of silk fibroin and polypyrrole (SP50) with a typical sponge-like porous structure and electrical conductivity similar to the native myocardium is developed. An electroactive engineered cardiac patch (SP50 ECP) with a certain thickness is constructed by applying electrical stimulation (ES) to the cardiomyocytes (CMs) on the scaffold. SP50 ECP can significantly express cardiac marker protein (α-actinin, Cx-43, and cTnT) and has better contractility and electrical coupling performance. Following in vivo transplantation, SP50 ECP shows a notable therapeutic effect in repairing infarcted myocardium. Not only can SP50 ECP effectively improves left ventricular remodeling and restore cardiac functions, such as ejection function (EF), but more importantly, improves the propagation of electrical pulses and promote the synchronous contraction of CMs in the scar area with normal myocardium, effectively reducing the susceptibility of MI rats to arrhythmias. In conclusion, this study demonstrates a facile approach to constructing electroactive ECPs based on porous conductive scaffolds and proves the therapeutic effects of ECPs in repairing the infarcted heart, which may represent a promising strategy for MI treatment.

A Matrix-Metalloproteinase-Responsive Hydrogel System for Modulating the Immune Microenvironment in Myocardial Infarction.

Injectable hydrogels carrying therapeutic factors to modulate the infarct immune microenvironment show great potential in the treatment of myocardial infarction (MI). However, conventional injectable hydrogels release therapeutic factors in an uncontrolled manner, which leads to poor treatment efficacy and acute side effects on normal tissues. In this work, a matrix metalloproteinase (MMP)2/9-responsive hydrogel system (MPGC4) is developed, considering the characteristics of the post-MI microenvironment. MPGC4 consists of tetra-poly(ethylene glycol) (PEG) hydrogels and a composite gene nanocarrier (CTL4) that is composed of carbon dots (CDots) coupled with interleukin-4 plasmid DNA via electrostatic interactions. MPGC4 can be automatically triggered to release CTL4 on demand after MI to regulate the infarct immune microenvironment. In addition, due to the photoluminescence properties of CDots, a large amount of viscoelastic MPGC4 is found to be retained in situ after injection into the infarct region without leakage. The in vitro results demonstrate that CTL4 promotes proinflammatory M1 macrophage polarization to the anti-inflammatory M2 subtype and contributes to cardiomyocyte survival through macrophage transition. In a rat model of MI, MPGC4 clears MMPs and precisely targets CTL4 to the infarcted region. In particular, MPGC4 improves cardiac function by modulating macrophage transition to reduce early inflammatory responses and proangiogenic activity.

Anti-inflammatory properties of S53P4 bioactive glass implant material.

OBJECTIVES: To assess whether the dissolution products of S53P4 bioactive glass (BG) affect cellular response of macrophages and clinically relevant peri­implant cell populations to dental implant particles in vitro. Cells chosen were human gingival fibroblasts (HGFs), osteoblasts and bone marrow derived stromal cells (HBMSCs). METHODS: Melt-derived S53P4 bioactive glass were prepared. HGFs, Saos-2 human osteoblastic cell line, HBMSCs and macrophages, derived from THP-1 human monocytic cell line, were cultured in the presence of particles from commercially pure titanium (Ti-CP4), grade 5 titanium alloy (Ti-6Al-4V), titanium-zirconium alloy (Ti-15Zr) or zirconia (Zr) (with respective diameters of 34.1 ± 3.8, 33.3 ± 4.4, 97.8 ± 8.2 and 71.3 ± 6.1 µm) with or without S53P4 dissolution products (conditioned media contained 327.30 ± 2.01 ppm Ca, 51.34 ± 0.41 ppm P and 61.48 ± 1.17 ppm Si, pH 8.01 ± 0.21). Inflammatory and macrophage polarisation markers including TNF-ɑ, IL-1, IL-6 and CD206 were quantified using enzyme-linked immunosorbent assay (ELISA). RESULTS: The presence of Ti-6Al-4V implant particles significantly induced the expression of pro-inflammatory markers in all tested cell types. S53P4 BG dissolution products regressed the particle induced up-regulation of pro-inflammatory markers and, appeared to suppress M1 macrophage polarisation. CONCLUSIONS: Implant particles, Ti-6Al-4V in particular, resulted in significant inflammatory responses from cells. S53P4 BG may possess anti-inflammatory properties and potentially mediate macrophage polarisation behaviour. CLINICAL SIGNIFICANCE: The findings highlight that the use and benefits of BG is a promising field of study. Authors believe more collective efforts are required to fully understand the reliability, efficiency and exact mechanisms of action of BG in the search for new generation of treatment modalities in dentistry.

Particle release from dental implants immediately after placement - An ex vivo comparison of different implant systems.

OBJECTIVES: Metallic element release during implant placement can lead to mucositis and peri-implantitis. Here, using ex vivo porcine mandibles, the release of metallic elements into the surrounding bone with different material and geometrical designs was quantified. METHODS: Implants from BioHorizons® and Straumann® (Bone level, tapered/cylindrical, 3/4 mm body diameter, Ti-CP4/Ti-6Al-4V/Ti-15Zr) systems were used. Micro computed tomography and inductively coupled plasma optical emission spectroscopy was used to visualise and quantify metallic elements in bone, following acid digestion. Implant surfaces were examined with scanning electron microscopy and internalization of implant particles by human gingival fibroblasts (HGFs) and RAW 264.7 macrophages were demonstrated in vitro. RESULTS: Implants with wider body diameters resulted in higher metallic element release. Ti-6Al-4V implants released significantly more metallic elements in comparison to both Ti-CP4 and Ti-15Zr devices with similar design and dimensions. Tapered Ti-CP4 implants released less compared to those with cylindrical design. Al three types of particles were internalized by HGFs and RAW 264.7. SIGNIFICANCE: Ti-CP4 and Ti-15Zr appear to be more suitable materials, however, further studies are required to elucidate the biological effects of the fine particles and/or metallic species from dental implants. Authors would like to raise the awareness in the dental profession community that careful evaluation of the materials used in dental implants and the potential risks of the individual constituents of any alloy are needed. The potential cytotoxicity of Ti-6Al-4V implant particles should be highlighted. Further investigations on the biological effect of the fine particles or metallic species released from dental implants are also needed.

Is titanium alloy Ti-6Al-4 V cytotoxic to gingival fibroblasts-A systematic review.

OBJECTIVES: Grade V titanium alloy (Ti-6Al-4 V) is a well-recognized metallic biomaterial for medical implants. There has been some controversy regarding the use of this alloy in medical devices in relation to the toxicity of vanadium. In Dentistry, Ti-6Al-4 V remains prevalent. This systematic review aims to evaluate the effects of Ti-6Al-4 V on cells relevant to oral environments such as gingival fibroblasts. MATERIALS AND METHODS: A literature search was undertaken for relevant English language publications in the following databases: Dental and Oral Science, Medline and Web of Science. The electronic search was supplemented with a search of references. RESULTS: After application of inclusion and exclusion criteria. A total of eight papers are included in this review. These papers were all in vitro studies and were categorized into whole implant, discs, or implant particles based on the type of test materials used in the studies. CONCLUSION: Based on the analyses of the eight included studies in this review, if Ti-6Al-4 V as a material is unchallenged, i.e., as a whole implant in pH neutral environments, there appears to be little effect on fibroblasts. If Ti-6Al-4 V is challenged through corrosion or wear (particle release), the subsequent release of vanadium and aluminium particles has an increased cytotoxic effect in vitro in comparison to commercially pure titanium, hence concerns should be raised in the clinical setting.

Particle release from implantoplasty of dental implants and impact on cells.

BACKGROUND: With increasing numbers of dental implants placed annually, complications such as peri-implantitis and the subsequent periprosthetic osteolysis are becoming a major concern. Implantoplasty, a commonly used treatment of peri-implantitis, aims to remove plaque from exposed implants and reduce future microbial adhesion and colonisation by mechanically modifying the implant surface topography, delaying re-infection/colonisation of the site. This in vitro study aims to investigate the release of particles from dental implants and their effects on human gingival fibroblasts (HGFs), following an in vitro mock implantoplasty procedure with a diamond burr. MATERIALS AND METHODS: Commercially available implants made from grade 4 (commercially pure, CP) titanium (G4) and grade 5 Ti-6Al-4 V titanium (G5) alloy implants were investigated. Implant particle compositions were quantified by inductively coupled plasma optical emission spectrometer (ICP-OES) following acid digestion. HGFs were cultured in presence of implant particles, and viability was determined using a metabolic activity assay. RESULTS: Microparticles and nanoparticles were released from both G4 and G5 implants following the mock implantoplasty procedure. A small amount of vanadium ions were released from G5 particles following immersion in both simulated body fluid and cell culture medium, resulting in significantly reduced viability of HGFs after 10 days of culture. CONCLUSION: There is a need for careful evaluation of the materials used in dental implants and the potential risks of the individual constituents of any alloy. The potential cytotoxicity of G5 titanium alloy particles should be considered when choosing a device for dental implants. Additionally, regardless of implant material, the implantoplasty procedure can release nanometre-sized particles, the full systemic effect of which is not fully understood. As such, authors do not recommend implantoplasty for the treatment of peri-implantitis.

Engineering an Artificial Membrane Vesicle Trafficking System (AMVTS) for the Excretion of ß-Carotene in Escherichia coli.

Large hydrophobic molecules, such as carotenoids, cannot be effectively excreted from cells by natural transportation systems. These products accumulate inside the cells and affect normal cellular physiological functions, which hinders further improvement of carotenoid production by microbial cell factories. In this study, we proposed to construct a novel artificial transport system utilizing membrane lipids to carry and transport hydrophobic molecules. Membrane lipids allow the physiological mechanism of membrane dispersion to be reconstructed and amplified to establish a novel artificial membrane vesicle transport system (AMVTS). Specifically, a few proteins in E. coli were reported or proposed to be related to the formation mechanism of outer membrane vesicles, and were individually knocked out or overexpressed to test their physiological functions. The effects on tolR and nlpI were the most significant. Knocking out both tolR and nlpI resulted in a 13.7% increase of secreted ß-carotene with a 35.6% increase of specific production. To supplement the loss of membrane components of the cells due to the increased membrane vesicle dispersion, the synthesis pathway of phosphatidylethanolamine was engineered. While overexpression of AccABCD and PlsBC in TW-013 led to 15% and 17% increases of secreted ß-carotene, respectively, the overexpression of both had a synergistic effect and caused a 53-fold increase of secreted ß-carotene, from 0.2 to 10.7 mg/g dry cell weight (DCW). At the same time, the specific production of ß-carotene increased from 6.9 to 21.9 mg/g DCW, a 3.2-fold increase. The AMVTS was also applied to a ß-carotene hyperproducing strain, CAR025, which led to a 24-fold increase of secreted ß-carotene, from 0.5 to 12.7 mg/g DCW, and a 61% increase of the specific production, from 27.7 to 44.8 mg/g DCW in shake flask fermentation. The AMVTS built in this study establishes a novel artificial transport mechanism different from natural protein-based cellular transport systems, which has great potential to be applied to various cell factories for the excretion of a wide range of hydrophobic compounds.

Polydimethylsiloxane (PDMS)-Based Flexible Optical Electrodes with Conductive Composite Hydrogels Integrated Probe for Optogenetics.

Optical electrodes are important tools for optogenetic research. Flexible optical electrodes represent a refinement over traditional fiber-based electrodes because they contact with target cells gently by reducing mechanical mismatch, thereby enhancing their long-term, stable signal acquisition capability. Until now, little attention has been paid to flexible intracortical optical electrodes. Here, we reported a novel flexible penetrating optical electrode with a probe made of composite hydrogels. We used polydimethylsiloxane (PDMS), a kind of transparent material, to fabricate waveguide by capillary assembly method with two tungsten wires inside providing mechanic support. Then one tungsten wire was withdrawn out and the microchannel was filled with hydrogel composed of polyvinyl alcohol (PVA), multi-walled carbon nanotubes (MWCNT), poly(3,4-ethylenedioxythiophene) (PEDOT), and polystyrene sulfonate (PSS) as an electrical recording and stimulation probe. With PDMS as the waveguide and PVA/MWCNT/PEDOT/PSS hydrogel as the electroprobe, the optical electrode becomes a flexible package. The morphology observed by scanning electron microscopy showed that the PVA/MWCNT/PEDOT/PSS hydrogel had a loose surface structure, which would allow the effective adhesion to target neurons. A buckling test showed that our electrode maintained bending strength comparable to that of previously reported flexible penetrating electrodes. Finally, the electrical properties showed a lower impedance and higher charge capacity after PEDOT/PSS modification. The flexible penetrating optical electrode we developed may be used for long-term in vivo optogenetics studies.

Biodegradable inorganic-organic hybrids of methacrylate star polymers for bone regeneration.

Hybrids that are molecular scale co-networks of organic and inorganic components are promising biomaterials, improving the brittleness of bioactive glass and the strength of polymers. Methacrylate polymers have high potential as the organic source for hybrids since they can be produced, through controlled polymerization, with sophisticated polymer architectures that can bond to silicate networks. Previous studies showed the mechanical properties of hybrids can be modified by polymer architecture and molar mass (MM). However, biodegradability is critical if hybrids are to be used as tissue engineering scaffolds, since the templates must be remodelled by host tissue. Degradation by-products have to either completely biodegrade or be excreted by the kidneys. Enzyme, or bio-degradation is preferred to hydrolysis by water uptake as it is expected to give a more controlled degradation rate. Here, branched and star shaped poly(methyl methacrylate-co-3-(trimethoxysilyl)propyl methacrylate) (poly(MMA-co-TMSPMA)) were synthesized with disulphide based dimethacrylate (DSDMA) as a biodegradable branching agent. Biodegradability was confirmed by exposing the copolymers to glutathione, a tripeptide which is known to cleave disulphide bonds. Cleaved parts of the star polymer from the hybrid system were detected after 2weeks of immersion in glutathione solution, and MM was under threshold of kidney filtration. The presence of the branching agent did not reduce the mechanical properties of the hybrids and bone progenitor cells attached on the hybrids in vitro. Incorporation of the DSDMA branching agent has opened more possibilities to design biodegradable methacrylate polymer based hybrids for regenerative medicine. STATEMENT OF SIGNIFICANCE: Bioactive glasses can regenerate bone but are brittle. Hybrids can overcome this problem as intimate interactions between glass and polymer creates synergetic properties. Implants have previously been made with synthetic polymers that degrade by water, however, they degrade catastrophically, causing rapid loss of strength. Polymers that degrade by biological agents may degrade at a more controlled rate, which should give time for tissue repair and transfer of load. Previously, hybrids made with star shaped poly(methyl methacrylate-co-3-(trimethoxysilyl)propyl methacrylate) (p(MMA-co-TMSPMA)) showed enhanced properties. However, methacrylates are not bio-degradable. Here, star shaped p(MMA-co-TMSPMA) was synthesized with a core that can be cleaved by glutathione, a tripeptide. On exposure to glutathione, the hybrid degraded, producing products with molecular weights below the kidney filtration threshold.

3D Printing of Biocompatible Supramolecular Polymers and their Composites.

A series of polymers capable of self-assembling into infinite networks via supramolecular interactions have been designed, synthesized, and characterized for use in 3D printing applications. The biocompatible polymers and their composites with silica nanoparticles were successfully utilized to deposit both simple cubic structures, as well as a more complex twisted pyramidal feature. The polymers were found to be not toxic to a chondrogenic cell line, according to ISO 10993-5 and 10993-12 standard tests and the cells attached to the supramolecular polymers as demonstrated by confocal microscopy. Silica nanoparticles were then dispersed within the polymer matrix, yielding a composite material which was optimized for inkjet printing. The hybrid material showed promise in preliminary tests to facilitate the 3D deposition of a more complex structure.

Nanofibrous poly(3-hydroxybutyrate)/poly(3-hydroxyoctanoate) scaffolds provide a functional microenvironment for cartilage repair.

Articular cartilage defects, when repaired ineffectively, often lead to further deterioration of the tissue, secondary osteoarthritis and, ultimately, joint replacement. Unfortunately, current surgical procedures are unable to restore normal cartilage function. Tissue engineering of cartilage provides promising strategies for the regeneration of damaged articular cartilage. As yet, there are still significant challenges that need to be overcome to match the long-term mechanical stability and durability of native cartilage. Using electrospinning of different blends of biodegradable poly(3-hydroxybutyrate)/poly(3-hydroxyoctanoate), we produced polymer scaffolds and optimised their structure, stiffness, degradation rates and biocompatibility. Scaffolds with a poly(3-hydroxybutyrate)/poly(3-hydroxyoctanoate) ratio of 1:0.25 exhibit randomly oriented fibres that closely mimic the collagen fibrillar meshwork of native cartilage and match the stiffness of native articular cartilage. Degradation of the scaffolds into products that could be easily removed from the body was indicated by changes in fibre structure, loss of molecular weight and a decrease in scaffold stiffness after one and four months. Histological and immunohistochemical analysis after three weeks of culture with human articular chondrocytes revealed a hyaline-like cartilage matrix. The ability to fine tune the ultrastructure and mechanical properties using different blends of poly(3-hydroxybutyrate)/poly(3-hydroxyoctanoate) allows to produce a cartilage repair kit for clinical use to reduce the risk of developing secondary osteoarthritis. We further suggest the development of a toolbox with tailor-made scaffolds for the repair of other tissues that require a 'guiding' structure to support the body's self-healing process.