Peptide-Based Biomaterials for Bone and Cartilage Regeneration.

The healing of osteochondral defects (OCDs) that result from injury, osteochondritis, or osteoarthritis and bear lesions in the cartilage and bone, pain, and loss of joint function in middle- and old-age individuals presents challenges to clinical practitioners because of non-regenerative cartilage and the limitations of current therapies. Bioactive peptide-based osteochondral (OC) tissue regeneration is becoming more popular because it does not have the immunogenicity, misfolding, or denaturation problems associated with original proteins. Periodically, reviews are published on the regeneration of bone and cartilage separately; however, none of them addressed the simultaneous healing of these tissues in the complicated heterogeneous environment of the osteochondral (OC) interface. As regulators of cell adhesion, proliferation, differentiation, angiogenesis, immunomodulation, and antibacterial activity, potential therapeutic strategies for OCDs utilizing bone and cartilage-specific peptides should be examined and investigated. The main goal of this review was to study how they contribute to the healing of OCDs, either alone or in conjunction with other peptides and biomaterials.

Recent Advances and Challenges in the Early Diagnosis and Treatment of Preterm Labor.

Preterm birth (PTB) is the primary cause of neonatal mortality and long-term disabilities. The unknown mechanism behind PTB makes diagnosis difficult, yet early detection is necessary for controlling and averting related consequences. The primary focus of this work is to provide an overview of the known risk factors associated with preterm labor and the conventional and advanced procedures for early detection of PTB, including multi-omics and artificial intelligence/machine learning (AI/ML)- based approaches. It also discusses the principles of detecting various proteomic biomarkers based on lateral flow immunoassay and microfluidic chips, along with the commercially available point-of-care testing (POCT) devices and associated challenges. After briefing the therapeutic and preventive measures of PTB, this review summarizes with an outlook.

Capra cartilage-derived peptide delivery via carbon nano-dots for cartilage regeneration.

Targeted delivery of site-specific therapeutic agents is an effective strategy for osteoarthritis treatment. The lack of blood vessels in cartilage makes it difficult to deliver therapeutic agents like peptides to the defect area. Therefore, nucleus-targeting zwitterionic carbon nano-dots (CDs) have immense potential as a delivery vehicle for effective peptide delivery to the cytoplasm as well as nucleus. In the present study, nucleus-targeting zwitterionic CDs have been synthesized as delivery vehicle for peptides while also working as nano-agents towards optical monitoring of cartilage healing. The functional groups of zwitterion CDs were introduced by a single-step microwave assisted oxidation procedure followed by COL II peptide conjugation derived from Capra auricular cartilage through NHS/EDC coupling. The peptide-conjugated CDs (PCDs) allows cytoplasmic uptake within a short period of time (∼30 m) followed by translocation to nucleus after ∼24 h. Moreover, multicolor fluorescence of PCDs improves (blue, green, and read channel) its sensitivity as an optical code providing a compelling solution towards enhanced non-invasive tracking system with multifunctional properties. The PCDs-based delivery system developed in this study has exhibited superior ability to induce ex-vivo chondrogenic differentiation of ADMSCs as compared to bare CDs. For assessment of cartilage regeneration potential, pluronic F-127 based PCDs hydrogel was injected to rabbit auricular cartilage defects and potential healing was observed after 60 days. Therefore, the results confirm that PCDs could be an ideal alternate for multimodal therapeutic agents.

Prefabricated 3D-Printed Tissue-Engineered Bone for Mandibular Reconstruction: A Preclinical Translational Study in Primate.

The advent of three dimensionally (3D) printed customized bone grafts using different biomaterials has enabled repairs of complex bone defects in various in vivo models. However, studies related to their clinical translations are truly limited. Herein, 3D printed poly(lactic-co-glycolic acid)/ß-tricalcium phosphate (PLGA/TCP) and TCP scaffolds with or without recombinant bone morphogenetic protein -2 (rhBMP-2) coating were utilized to repair primate's large-volume mandibular defects and compared efficacy of prefabricated tissue-engineered bone (PTEB) over direct implantation (without prefabrication). 18F-FDG PET/CT was explored for real-time monitoring of bone regeneration and vascularization. After 3-month's prefabrication, the original 3D-architecture of the PLGA/TCP-BMP scaffold was found to be completely lost, while it was properly maintained in TCP-BMP scaffolds. Besides, there was a remarkable decrease in the PLGA/TCP-BMP scaffold density and increase in TCP-BMP scaffolds density during ectopic (within latissimus dorsi muscle) and orthotopic (within mandibular defect) implantation, indicating regular bone formation with TCP-BMP scaffolds. Notably, PTEB based on TCP-BMP scaffold was successfully fabricated with pronounced effects on bone regeneration and vascularization based on radiographic, 18F-FDG PET/CT, and histological evaluation, suggesting a promising approach toward clinical translation.

Bioinspired channeled, rhBMP-2-coated ß-TCP scaffolds with embedded autologous vascular bundles for increased vascularization and osteogenesis of prefabricated tissue-engineered bone.

To date, the recovery of large bone defects is a major clinical challenge despite the availability of numerous therapeutic procedures including tissue engineering. Although there is a pressing need for large tissue-engineered constructs, inadequate vascularization remains an insurmountable barrier for successful clinical translation. Considering that vascularization is a prerequisite for osteogenesis, we proposed an advanced design of large customized porous ß-tricalcium phosphate (TCP) scaffolds with biomimetic vascular hierarchy which upon embedding of femoral axial vascular bundles significantly improved overall vascularity of the scaffolds. Such scaffolds also promoted osteogenesis when they were coated with recombinant bone morphogenetic protein-2 (rhBMP-2). Compared to the conventional TCP scaffolds (S), the newly designed multi-channeled ß-TCP (CS) scaffolds led to adequate blood vessels and bone-like tissue formation throughout their porous hierarchy within 4 weeks of implantation. Especially, the scaffolds coated with rhBMP-2 and embedded with flow-through vascular bundle (FVB) were able to form more uniform vascularized bone within 2 weeks post-implantation. Based on the clinical, radiographic, angiographic and histological assessments, the newly designed multi-channeled scaffolds were found to be promising for successful recovery of large bone defects.

pH-labile and photochemically cross-linkable polymer vesicles from coumarin based random copolymer for cancer therapy.

The stability of a drug payload inside a nanocarrier at physiological environment and the release of the said drug at specific tumor cells in a sustainable manner are the two most important factors that determine the efficiency of a smart targeted drug-delivery system. In this work, 2-hydroxyethyl methacrylate and a coumarin-based methacrylate monomer containing ß-thiopropionate moiety were copolymerized via reversible addition-fragmentation chain transfer (RAFT) process, followed by characterization using NMR and GPC. The said copolymer self-assembled at physiological pH to form vesicular nano-aggregates which was confirmed using DLS, TEM and by fluorescence measurements. These vesicles were further stabilized by photochemical crosslinking via coumarin (2π + 2π) cycloaddition reaction. These cross-linked vesicles (CVs) exhibited a 38% reduction in premature drug leakage as compared to the uncross-linked vesicles (UCVs) at physiological pH. Additionally, a slow hydrolysis of the ß-thiopropionate moieties under mildly acidic conditions prevalent in tumor cells resulted in disassembly of the vesicles, thereby releasing the loaded drug in a sustainable manner. Studies related to in vitro toxicity, efficiency of cellular uptake and pH-responsive antineoplastic activity of doxorubicin (DOX) loaded in the cross-linked vesicles (CVs) toward cancer cell lines were undertaken. A significant reduction in IC50 was noticed for DOX-loaded CVs in comparison to free DOX toward MG63 cancer cell lines, making these vesicles as potent nanocarrier systems for cancer therapy.

Isolation and mass spectrometry based hydroxyproline mapping of type II collagen derived from Capra hircus ear cartilage.

Collagen II (COLII), the most abundant protein in vertebrates, helps maintain the structural and functional integrity of cartilage. Delivery of COLII from animal sources could improve cartilage regeneration therapies. Here we show that COLII can be purified from the Capra ear cartilage, a commonly available bio-waste product, with a high yield. MALDI-MS/MS analysis evidenced post-translational modifications of the signature triplet, Glycine-Proline-Hydroxyproline (G-P-Hyp), in alpha chain of isolated COLII (COLIIA1). Additionally, thirty-two peptides containing 59 Hyp residues and a few G-X-Y triplets with positional alterations of Hyp in COLIIA1 are also identified. Furthermore, we show that an injectable hydrogel formulation containing the isolated COLII facilitates chondrogenic differentiation towards cartilage regeneration. These findings show that COLII can be isolated from Capra ear cartilage and that positional alteration of Hyp in its structural motif, as detected by newly developed mass spectrometric method, might be an early marker of cartilage disorder.

Osteochondral Defects Healing Using Extracellular Matrix Mimetic Phosphate/Sulfate Decorated GAGs-Agarose Gel and Quantitative Micro-CT Evaluation.

Tissue engineering has a major emphasis in creating tissue specific extracellular ambiance by altering chemical functionalities of scaffold materials. Heterogeneity of osteochondral tissue necessitates tailorable bone and cartilage specific extracellular environment. Carboxylate- and sulfate-functionalized glycosaminoglycans (GAGs) in cartilage extracellular matrix (ECM) create an acidic ambience to support chondrogenic activity, whereas phosphate-rich environment in bone enables chelation of calcium leading to the formation of mineralized matrix along with an alkaline environment to support osteogenesis. In this study, chitosan, a naturally occurring GAGs, was functionalized with phosphate/sulfate groups analogous to bone/cartilage ECM and incorporated in thermogelling agarose hydrogel for delivery to osteochondral defects. In vitro studies revealed significantly higher adhesion and proliferation of adipose derived mesenchymal stem cells (ADMSCs) with blended hydrogels as compared to that of native agarose. Cell differentiation and RT-PCR studies of the phosphorylated hydrogels revealed higher osteogenic potential, while sulfated hydrogels demonstrated enhanced chondrogenic activity in comparison to agarose. Recovery of osteochondral defects after delivery of the thermoresponsive agarose-based hydrogels decorated with phosphorylated derivatives showed significantly higher bone formation. On the other hand, cartilage formation was significant with chitosan sulfate decorated hydrogels. The study highlights the role of chitosan derivatives in osteochondral defect healing, especially phosphorylated ones as bone promoter, whereas sulfated ones act as cartilage enhancer, which was quantitatively distinguished through micro-CT-based noninvasive imaging and analysis.

Simultaneous hydrothermal bioactivation with nano-topographic modulation of porous titanium alloys towards enhanced osteogenic and antimicrobial responses.

Post-implantation failure associated with insufficient host tissue integration at the bone-implant interface and aseptic loosening is a major concern in orthopaedics as well as in dentistry. To overcome the failure in early stages of implantation, prosthetic design combining the mechanisms of porosity guided bone ingrowth along with topographic manipulation of osteogenic cells over bacterial colonization would be an ideal choice, although achieving such a goal is highly challenging. In this study, facile rapid hydrothermal synthesis of nanostructures with simultaneous deposition of hydroxyapatite on the titanium alloy surface was demonstrated by using an aqueous sodium tripolyphosphate and calcium hydroxide mixture. Nanostructures with wire-like morphology exhibited significantly higher osteogenic related gene expression (COL I, OPN, and OCN) through differentiation of adipose derived mesenchymal stem cells as well as the bactericidal response against S. aureus and E. coli as compared to other nanotopographic features. The same also exhibited elongated cell morphology with the highest expression of paxillin towards cell boundaries as compared to the polished surface with flattened cell morphology and localized expression of paxillin around the nucleus. Implantation of treated porous Ti6Al4V samples representing a multiscalar hierarchy with wire-like nanostructures accelerated osteochondral healing in rabbits without any major signs of infection. Also, significantly higher bone formation was observed within the defects implanted with treated porous Ti6Al4V (44.0%) as compared to that of untreated porous samples (36.9%) as well as empty defects (19.6%).

Influence of Porosity and Pore-Size Distribution in Ti6Al4 V Foam on Physicomechanical Properties, Osteogenesis, and Quantitative Validation of Bone Ingrowth by Micro-Computed Tomography.

Cementless fixation for orthopedic implants aims to obviate challenges associated with bone cement, providing long-term stability of bone prostheses after implantation. The application of porous titanium and its alloy-based implants is emerging for load-bearing applications due to their high specific strength, low stiffness, corrosion resistance, and superior osteoconductivity. In this study, coagulant-assisted foaming was utilized for the fabrication of porous Ti6Al4 V using egg-white foam. Samples with three different porosities of 68.3%, 75.4%, and 83.1% and average pore sizes of 92, 178, and 297 µm, respectively, were prepared and subsequently characterized for mechanical properties, osteogenesis, and tissue ingrowth. A microstructure-mechanical properties relationship study revealed that an increase of porosity from 68.3 to 83.1% increased the average pore size from 92 to 297 µm with the subsequent reduction of compresive strength by 85% and modulus by 90%. Samples with 75.4% porosity and a 178 µm average pore size produced signifcant osteogenic effects on human mesenchymal stem cells, which was further supported by immunocytochemistry and real-time polymerase chain reaction data. Quantitative assessment of bone ingrowth by micro-computed tomography revealed that there was an approximately 52% higher bone formation and more than 90% higher bone penetration at the center of femoral defects in rabbit when implanted with Ti6Al4 V foam (75.4% porosity) compared to the empty defects after 12 weeks. Hematoxylin and eosin (H&E) and Masson trichrome (MT) staining along with energy-dispersive X-ray mapping on the sections obtained from the retrieved bone samples support bone ingrowth into the implanted region.

Investigating the potential of human placenta-derived extracellular matrix sponges coupled with amniotic membrane-derived stem cells for osteochondral tissue engineering.

Osteochondral injuries are challenging to repair due to their complex tissue anatomy and restricted self-repairing ability associated with a limited blood supply. Osteochondral tissue engineering is an important clinical aspect of the management and treatment of cartilage and underlying bone. In the present study, we fabricated human placenta-derived extracellular matrix sponges (PEMS) for repair of osteochondral tissue through a decellularization process. There were no significant cellular components present in the PEMS; hematoxylin & eosin/DAPI staining, DNA quantification and agarose gel electrophoresis were used to evaluate the extent of decellularization. Moreover, no significant alteration to the collagen and glycosaminoglycan (native extracellular matrix) content of the PEMS was observed. PEMS in vitro provided a non-cytotoxic environment rich in bioactive cues for human amniotic membrane-derived stem cells (HAMSCs) to proliferate in and differentiate into chondrogenic and osteogenic lineages under induction. Histological analysis at 28 days after the PEMS were subcutaneously implanted demonstrated no severe immune response in the host and supported the formation of blood vessels. To assess the osteochondral tissue repair ability of PEMS, cell-free PEMS (CFP) and cell-seeded PEMS (CSP) were implanted at osteochondral defect sites in a rabbit model. Histological scores indicated that osteochondral regeneration was more successful in the defects filled with CSP compared to those filled with CFP and empty defects (ED) after 60 days of implantation. In summary, a naturally derived biocompatible scaffold composed of extracellular matrix from human placenta has been successfully developed for osteochondral tissue engineering.