Supportive membranes have recently been applied to treat periodontal disease in order to achieve periodontal tissue regeneration. The crucial role of these membranes is to facilitate the restoration of the structural and functional periodontium. Bovine pericardium (BP) is mainly composed of collagen type I, which was demonstrated to have good mechanical properties and biological regenerative potential. Our research aimed to extend the application of membrane derived from BP to periodontal disease treatment. However, the fabrication method to achieve a xenogenic-free membrane with the mechanical properties required for periodontal treatment is rarely mentioned. Therefore, a procedure for the extraction and modification of BP using sodium dodecyl sulfate (SDS) and glutaraldehyde (GA) was developed. BP was harvested and decellularized using different SDS concentrations (0.05–0.3%). GA was used to further modify the membranes to achieve suitable thickness, mechanical strength, and pore size. A combination protocol of 0.15% SDS treatment for 12 h with continuous agitation combined with 0.1% GA for 6 h for membrane fabricating was applied. The modified BP (mBP) had the targeted characteristics, such as 0.2–0.5 mm thickness, approximately 10 MPa in tensile strength, 30% in strain force, and a pore size <5 µm, which is comparable to commercially available collagen membranes. Findings from this study demonstrated that the established method was effective in preparing BP membrane for periodontal treatment while decreasing the concentration of reagents and processing time. Moreover, our modified membrane was found to have no cytotoxicity but supports the migration, attachment, and proliferation of human gingival fibroblasts in vitro. Taken together, these results confirmed that mBP is suitable for application in periodontal disease treatment and regeneration.

(1) Background: Hemocompatibility is a critical challenge for tissue-derived biomaterial when directly contacting the bloodstream. In addition to surface modification with heparin, endothelialization of the grafted material is suggested to improve long-term clinical efficacy. This study aimed to evaluate the ability to endothelialize in vitro of heparinized bovine pericardial scaffolds. (2) Methods: bovine pericardial scaffolds were fabricated and heparinized using a layer-by-layer assembly technique. The heparinized scaffolds were characterized for heparin content, surface morphology, and blood compatibility. Liquid extraction of the samples was prepared for cytotoxicity testing on human endothelial cells. The in-vitro endothelialization was determined via human endothelial cell attachment and proliferation on the scaffold. (3) Results: The heparinized bovine pericardial scaffold exhibited a heparin coating within its microfiber network. The scaffold surface immobilized with heparin performed good anti-thrombosis and prevented platelet adherence. The proper cytotoxicity impact was observed for a freshly used heparinized sample. After 24 h washing in PBS 1X, the cell compatibility of the heparinized scaffolds was improved. In-vitro examination results exhibited human endothelial cell attachment and proliferation for 7 days of culture. (4) Conclusions: Our in-vitro analysis provided evidence for the scaffold’s ability to support endothelialization, which benefits long-term thromboresistance.

Bovine pericardium has been proposed as an available material for tissue engineering and bioprosthetic reconstruction. In this study, bovine pericardium was fabricated into a scaffold for culturing and chondrogenic differentiation of human adipose-derived stem cells (hADSCs). Bovine pericardium was treated in 10 mM Tris-HCl and 0.15% SDS, followed by crosslinking in 0.1% glutaraldehyde. Treated bovine pericardium (tBP) was characterized as a slight yellowish thin membrane with enhanced tensile strength and strain property. The membrane maintained stability under enzymatic conditions for up to 16 days of incubation. The results confirmed tBP as a cell-friendly scaffold for hADSCs due to low cytotoxicity and its ability to support an appropriate attachment and proliferation of hADSCs. Moreover, there was an accumulation of the extracellular matrix proteoglycan in tBP seeded with hADSCs after 7 and 14 days of chondrogenic induction. COMP as a specific marker of chondrogenesis was detected after 7 days, whereas type X-a1 collagen (Col10a1) expression was stable up to day 14. However, minor expression of aggrecan was found. Taken together, these results indicate that tBP is a potential scaffold for hADSCs for cartilage tissue engineering.Key words: Bovine pericardium, scaffold, adipose-derived stem cells, chondrogenic differentiation, cartilage regeneration, augmentation rhinoplasty.

Incorporation of extracellular matrix (ECM) components into bioinks can enhance biological functionality but often compromises print fidelity and structural stability. This study developed a dual-stage calcium chloride (CaCl₂) crosslinking strategy to incorporate Wharton’s jelly-derived ECM (WJ-ECM) into a gelatin-alginate bioink for bioprinted scaffold fabrication. A baseline formulation (BGA) and a WJ-ECM-supplemented formulation (BGAE, 1 mg/mL) were pre-crosslinked with 14 mM CaCl₂ prior to extrusion, followed by secondary crosslinking in 0.5 M CaCl₂ post-printing. Both formulations exhibited comparable viscosity (20–180 kcP) and high print fidelity (Pr = 0.99 ± 0.01 for BGA; 0.95 ± 0.01 for BGAE), with scaffolds displaying well-defined architecture and over 84% of pores within the target range (160–270 µm). FTIR analysis confirmed WJ-ECM incorporation without detectable alteration of the primary matrix structure. Both scaffolds were non-cytotoxic and supported fibroblast viability; BGAE constructs showed greater cell coverage over 14 days when surface-seeded and more stable fluorescence intensity through 28 days when encapsulated. In a murine thermal burn model, BGAE-treated wounds demonstrated more advanced re-epithelialization and more continuous epidermal coverage at day 14 compared to controls. These findings indicate that dual-stage crosslinking enables WJ-ECM integration while preserving printability, offering a practical platform for bioactive skin tissue engineering applications.

Collagen membrane that was mainly from the bovine and porcine pericardium has been studied and widely applied for most manufacturers of bioprothetic materials, especially periodontal therapeutics. As many studied, bovine pericardium has higher collagen content than that from porcine, although both membranes have no significant different in calcification when treated with glutaraldehyde. In this study, acellular bovine was used to fabricated a biomembrane for periodontal reconstruction and regeneration. Acellular bovine pericardium was treated with glutaraldehyde at different concentrations and time points. The treatment of 0,1% glutaraldehyde for 6 hours was determined as the optimal fabrication protocol according to membrane properties, including thickness (0.2mm-0.5mm), tensile strength (5MPa-12MPa), tensile strain (15%-50%), around 5µm pore size, limited in vitro degradation (1%-5%), which met all criteria parameters for periodontal treatment. The fabricated membrane presented in vitro biocompatibility to human fibroblasts according to ISO10993-5. Moreover, the membrane promoted the in vitro migration of humane gingival fibroblasts, which indicated the application as a guided biomembrane in periodontal reconstuction and regeneration.