Enzymatic synthesis of furan-based copolymers: Material characterization and potential for biomedical applications.

BACKGROUND: Today's growing demand for advanced and sustainable polyester materials is driven by an increasing awareness of the environmental impact of traditional materials, emphasizing the need for eco-friendly alternatives. Sustainability has become central in materials development, including the biomedical area, where biobased and environmentally friendly solutions are a rapidly growing field. OBJECTIVES: This research aims to comprehensively evaluate a new enzymatically catalyzed furan-based copolymer, poly(decamethylene furanoate)-co-(dilinoleic furanoate) (PDF-DLF), with a 70-30 wt% hard-to-soft segment ratio. Then, its performance across medical applications is explored, with a particular focus on its potential as a nanofibrous scaffolding material. MATERIAL AND METHODS: PDF-DLF was synthesized from biobased monomers using Candida antarctica lipase B (CAL-B) as the biocatalyst. Material characterization included dynamic mechan­ical thermal analysis (DMTA) to assess the mechanical behavior and thermal properties. Enzymatic degradation studies determined biodegradability, while cytotoxicity tests established in vitro biocompatibility. The copolymer was electrospun into nanofibers, with scanning electron microscopy (SEM) employed to analyze their morphology. RESULTS: PDF-DLF displays mechanical and thermal properties indicating high storage modulus and 2 main temperature transitions. Enzymatic degradation studies and cytotoxicity assessments confirm biodegradability and in vitro biocompatibility. Electrospinning successfully transformed the copolymer into nanofibers with diameters ranging from 500 nm to 700 nm. CONCLUSIONS: This study significantly advances our understanding of sustainable polyesters with versatile processing capabilities. The successful electrospinning highlights its potential as a biodegradable scaffold for medical engineering, supported by biocompatibility and sufficient mechanical properties. It opens new opportunities for sustainable materials in critical biomedical industries, including tissue engineering.

Bone repair potential of collagen-poly(3-hydroxybutyrate)-carbon nanotubes scaffold loaded with mesenchymal stem cells for the reconstruction of critical-sized mandibular defects.

The poor structural stability of collagen (COL) upon hydration poses a significant challenge in tissue engineering (TE). To overcome this limitation, the incorporation of hydrophobic polymers such as poly(3-hydroxybutyrate) (PHB), and nanomaterials such as carbon nanotubes (CNTs) has been explored. In this study, we investigated the physical, chemical, and biological characteristics of COL-based scaffolds modified with PHB and CNTs for bone tissue engineering (BTE) applications. The tensile strength analysis revealed a substantial improvement in the ultimate tensile strength with the addition of 10 % PHB and 4 % CNTs. Scanning electron microscopy (SEM) images depicted a denser and more compact structure resulting from the presence of PHB and CNTs, enhancing the scaffold's mechanical properties. Fourier-transform infrared spectroscopy (FTIR) confirmed the successful incorporation of PHB and CNTs into the composite scaffold, maintaining the chemical integrity of COL. Stereological studies also conducted in a rat model with induced critical-sized bone defects in the mandibular bone further emphasize the substantial increase in bone formation and reduction in defect volume achieved by the scaffold loaded with stem cells. These findings underscore the promising approach to enhance bone healing, using COL-based scaffolds loaded with stem cells, and the favorable results obtained in this study can contribute to the advancement of BTE strategies.

Comparison effect of collagen/P3HB composite scaffold and human amniotic membrane loaded with mesenchymal stem cells on colon anastomosis healing in male rats.

Covering surgical wounds with biomaterials, biologic scaffolds, and mesenchymal stem cells (MSCs) improves the healing process and reduces postoperative complications. This study was designed to evaluate and compare the effect of MSC-free/MSC-seeded new collagen/poly(3-hydroxybutyrate) (COL/P3HB) composite scaffold and human amniotic membrane (HAM) on the colon anastomosis healing process. COL/P3HB scaffold was prepared using freeze-drying method. MSCs were isolated and characterized from rat adipose tissue. After biocompatibility evaluation by MTT assay, MSCs were seeded on the scaffold and HAM by micro-mass seeding technique. In total, 35 male rats were randomly divided into five groups. After the surgical procedure, cecum incisions were covered by the MSC-free/MSC-seeded scaffold or HAM. Incisions in the control group were only sutured. One month later, the healing process was determined by stereological analysis. The Kruskal-Wallis followed by Dunn's tests were utilized for statistical outcome analysis (SPSS software version 21). COL/10% P3HB scaffold showed the best mechanical and structural properties (7.86 MPa strength, porosity more than 75%). MTT assay indicated that scaffold and especially HAM have suitable biocompatibility. Collagenization and neovascularization were significantly higher, and necrosis was considerably lower in all treated groups in comparison with the controls. MSC-seeded scaffold and HAM significantly decrease inflammation and increase gland volume compared with other groups. The MSC-seeded HAM was significantly successful in decreasing edema compared with other groups. Newly synthesized COL/P3HB scaffold improves the colon anastomosis healing; however, the major positive effect belonged to HAM. MSCs remarkably increase their healing process. Further investigations may contribute to confirming these results in other wound healing.

Attenuation of osteoarthritis progression through intra-articular injection of a combination of synovial membrane-derived MSCs (SMMSCs), platelet-rich plasma (PRP) and conditioned medium (secretome).

PURPOSE: Osteoarthritis (OA) as a progressive destructive disease of articular cartilage is the most common joint disease characterized by reduction of joint cartilage thickness, demolition of cartilage surface and new bone formation. To overcome these problems, the purpose of the current research was to evaluate and compare the in vivo effects of synovial membrane-derived mesenchymal stem cell (SMMSCs), platelet-rich plasma (PRP) and conditioned medium (secretome) on collagenase II-induced rat knee osteoarthritis (KOA) remedy. METHODS: For the first step, SMMSCs were isolated and characterized. Also, secretome was collected from SMMSCs culture. Furthermore, PRP was collect from the rat heart venous blood. Second, two injection of collagenase II with an interval of 3 days was performed in the knee intra-articular space to induce osteoarthritis. Two weeks later, animals were randomly divided into 6 groups. Control group without treatment, positive group: taken an intra-articular sodium hyaluronate injection (0.1 ml), treatment groups taken an intra-articular injection of; treatment 1: SMMSCs (5 × 106), treatment 2: SMMSCs (5 × 106)/secretome (50 µl), treatment 3: SMMSCs (5 × 106)/PRP (50 µl), and treatment 4: SMMSCs (5 × 106)/ secretome (50 µl)/ PRP (50 µl). Three months later, rats were killed and the following assessments were executed: radiography, histopathology, and immunohistochemistry. RESULTS: Our findings represented that a combination of the SMMSCs/secretome/PRP had a considerable effect on glycosaminoglycans (GAGs) and collagen II contents, articular cartilage preservation, compared with other groups. In addition, combination of the SMMSCs with PRP and secretome showed the lowest expression of mmp3, while SOX9 had the highest expression in comparison with other groups. Also, SMMSCs-injected groups demonstrated better results compared with positive and control groups. CONCLUSIONS: Injecting a combination of the SMMSCs/secretome/PRP resulted in better efficacy in terms of joint space width, articular cartilage surface continuity and integrity, sub-chondral bone and ECM constituents such as collagen II. Indeed, transplantation of this combination could be considered as a preliminary therapy for clinical trial study in the future.

Synthesis of Hydrophilic Poly(butylene succinate-butylene dilinoleate) (PBS-DLS) Copolymers Containing Poly(Ethylene Glycol) (PEG) of Variable Molecular Weights.

Polymeric materials have numerous applications from the industrial to medical fields because of their vast controllable properties. In this study, we aimed to synthesize series of poly(butylene succinate-dilinoleic succinate-ethylene glycol succinate) (PBS-DLS-PEG) copolymers, by two-step polycondensation using a heterogeneous catalyst and a two-step process. PEG of different molecular weights, namely, 1000 g/mol and 6000 g/mol, was used in order to study its effect on the surface and thermal properties. The amount of the PBS hard segment in all copolymers was fixed at 70 wt%, while different ratios between the soft segments (DLS and PEG) were applied. The chemical structure of PBS-DLS-PEG was evaluated using Fourier transform infrared spectroscopy and nuclear magnetic resonance spectroscopy. Gel permeation chromatography was used to determine the molecular weight and dispersity index. The results of structural analysis indicate the incorporation of PEG in the macrochain. The physical and thermal properties of the newly synthesized copolymers were also evaluated using water contact angle measurements, differential scanning calorimetry and dynamic thermomechanical analysis. It was found that increasing the amount of PEG of a higher molecular weight increased the surface wettability of the new materials while maintaining their thermal properties. Importantly, the two-step melt polycondensation allowed a direct fabrication of a polymeric filament with a well-controlled diameter directly from the reactor. The obtained results clearly show that the use of two-step polycondensation in the melt allows obtaining novel PBS-DLS-PEG copolymers and creates new opportunities for the controlled processing of these hydrophilic and thermally stable copolymers for 3D printing technology, which is increasingly used in medical techniques.

Poly(3-Hydroxybutyrate)-Multiwalled Carbon Nanotubes Electrospun Scaffolds Modified with Curcumin.

Appropriate selection of suitable materials and methods is essential for scaffolds fabrication in tissue engineering. The major challenge is to mimic the structure and functions of the extracellular matrix (ECM) of the native tissues. In this study, an optimized 3D structure containing poly(3-hydroxybutyrate) (P3HB), multiwalled carbon nanotubes (MCNTs) and curcumin (CUR) was created by electrospinning a novel biomimetic scaffold. CUR, a natural anti-inflammatory compound, has been selected as a bioactive component to increase the biocompatibility and reduce the potential inflammatory reaction of electrospun scaffolds. The presence of CUR in electrospun scaffolds was confirmed by 1H NMR and Fourier-transform infrared spectroscopy (FTIR). Scanning electron microscopy (SEM) revealed highly interconnected porosity of the obtained 3D structures. Addition of up to 20 wt% CUR has enhanced mechanical properties of the scaffolds. CUR has also promoted in vitro bioactivity and hydrolytic degradation of the electrospun nanofibers. The developed P3HB-MCNT composite scaffolds containing 20 wt% of CUR revealed excellent in vitro cytocompatibility using mesenchymal stem cells and in vivo biocompatibility in rat animal model study. Importantly, the reduced inflammatory reaction in the rat model after 8 weeks of implantation has also been observed for scaffolds modified with CUR. Overall, newly developed P3HB-MCNTs-CUR electrospun scaffolds have demonstrated their high potential for tissue engineering applications.

Crosslinked porous three-dimensional cellulose nanofibers-gelatine biocomposite scaffolds for tissue regeneration.

Gelatine is a biocompatible and natural polymer with chemical properties similar to the extracellular matrix. However, it has poor mechanical properties and sensitive to enzymatic biodegradation that limits its application in 3D scaffold fabrication. Cellulose nanofibrous (CNF) offers biocompatibility, high surface area and excellent mechanical properties with slow in-vivo degradation. To fine tune their properties, CNF, and gelatine (CNF-GEL) were blended to form biocomposite aerogels. Epichlorohydrin (EPH) was incorporated into CNF-GEL as a chemical crosslinker to investigate its effect on the physiochemical, mechanical, and biological properties of the biocomposite aerogels both in-vitro and in-vivo. Regardless of the composition of the prepared aerogels, they possessed porosity of >90% with the pore size of 7-135 µm, which was confirmed in the morphological analysis. The presence of EPH improved the chemical interaction between CNF and gelatine, hence enhanced the compressive strength compared to uncrosslinked samples. The formulation of crosslinked CNF-GEL 90:10 offered the highest compressive strength of 61.35 kPa. The in-vitro and in-vivo studies showed adequate cytocompatibility, cell viability and cell attachment in the optimal crosslinked formulation with tuned enzymatic degradation. Antimicrobial property was also achieved in the optimal scaffold by incorporating curcumin as an antimicrobial agent.

In Vitro and In Vivo Evaluation of Poly (3-hydroxybutyrate)/Carbon Nanotubes Electrospun Scaffolds for Periodontal Ligament Tissue Engineering.

STATEMENT OF THE PROBLEM: Tissue engineering was an idea, today it has become a potential therapy for several tissues in dentistry, such as periodontal disease and oral mucosa. PURPOSE: In this experimental study, periodontal regeneration is one of the earliest clinical disciplines that has achieved therapeutic application in tissue engineering. The aim of the present study was to prepare electrospun Poly (3-hydroxybutyrate) (PHB)/1% Carbon nanotubes (CNTs) scaffolds for periodontal regeneration. MATERIALS AND METHOD: 1% w/v of CNTs was added to the polymer solutions and electrospinned. Physical properties of the scaffolds were evaluated by scanning electron microscopy (SEM) and universal testing machine. Chemical characterization of the scaffolds was also assessed by Fourier-transform infrared spectroscopy (FTIR). Biological properties of the scaffolds were also evaluated in vitro by culturing periodontal ligament stem cells (PDLSCs) on the scaffolds for 10 days and in vivo by Implanting the scaffolds in rat model for 5 weeks. RESULTS: Results showed that the scaffolds mimicked fibrous connective tissue of the (PDL). CNTs improved the mechanical properties, similar to 23-55 years old human PDL. In vitro biocompatibility study showed more attachment and proliferation of the PDLSCs for PHB/1%CNTs scaffolds compared to the PHB controls. In vivo study showed that CNTs in the scaffolds caused mild foreign body type giant cell reaction, moderate vascularization, and mild inflammation. CONCLUSION: The results showed that the PHB/1%CNTs composite scaffolds might be potentially useful in periodontal regeneration.

The effects of adipose tissue-derived stem cells seeded onto the curcumin-loaded collagen scaffold in healing of experimentally- induced oral mucosal ulcers in rat.

OBJECTIVES: Various therapeutic approaches, including stem-cell-based strategies and tissue engineering, have been proposed for oral ulcerative lesions. We investigated the effects of adipose tissue-derived stem cells (ADSCs) seeded onto the curcumin-loaded collagen scaffold in the mucosal healing of oral ulcers in rats. MATERIALS AND METHODS: The current experimental study was conducted on 40 male Sprague-Dawley rats. Oral ulcers were created over both sides of buccal mucosa, and the rats were randomly divided into four equal groups: 1) an untreated group (negative control); 2) Teriadent-treated group (positive control); 3) group treated with curcumin-loaded collagen scaffold; and 4) group received the ADSCs (3 × 106 cells) seeded onto the curcumin-loaded collagen scaffold. Rats were sacrificed on 3rd and 7th day after ulceration for histopathological examination as well as measurement of tissue levels of myeloperoxidase (MPO), superoxide dismutase (SOD), and Interleukin-1 beta (IL-1ß) activity. RESULTS: Compared with the negative control, the tissue levels of MPO and IL-1ß were significantly decreased in all treated groups (P<0.0001); however, the SOD activity was elevated (P<0.0001). The highest SOD activity as well as the lowest MPO and IL-1ß levels were observed in the ADSCs-curcumin-loaded collagen scaffold group. The ulcer healing process at 3rd and 7th day follow-up was much more progressed in the ADSCs-curcumin-loaded collagen scaffold group in comparison with the untreated group (P=0.037 and P=0.004, respectively). CONCLUSION: According to the findings of this study, ADSCs seeded onto the curcumin-loaded collagen scaffold seems to have a promising potential for oral ulcer healing applications.