In vivo study of bovine hydroxyapatite-gelatin-hydroxypropyl methylcellulose with alendronate as injectable bone substitute composite in osteoporotic animal model.

The injectable bone substitute (IBS) is a self-setting local drug delivery system that adjusts the shape of the bone gap in the fracture. This study aimed to examine the effectiveness of IBS composites of bovine hydroxyapatite (BHA) and alendronate (Ale) in accelerating bone growth in osteoporotic rats. IBS was made by mixing BHA with gelatin 5%, hydroxypropyl methylcellulose (HPMC) 2%, and Ale 10%. The physical properties of IBS were viscosity, injectability, and density tests. Twenty-four female Wistar rats were divided into four groups. After 8 weeks, 2 mm gap was made in the right femur of all rats and filled with IBS. The healing process was observed after 6 weeks with X-ray imaging and H and E staining. The obtained results showed viscosity, injectability, and density value of IBS from 30.4 to 39.4 dPa.s, 98.22%-98.64%, and 0.6325-0.8409 g/cm3, respectively. X-ray imaging and histology results proved the condition of osteoporosis in rats with ovariectomy. The addition of BHA-Gel-HPMC-Ale significantly affected the number of osteoblasts, osteocytes, and osteoclasts (P < 0.05). After 45 days of observation, the addition of BHA-Gel-HPMC-Ale showed the highest mean number of osteoblasts, osteocytes, and osteoclasts, which were 25.00 ± 3.00, 64.33 ± 11.15, and 5.67 ± 0.58 compared to BHA-Gel-HPMC and positive control groups. The BHA-Gel-HPMC-Ale IBS has the potential to reverse osteoporosis. Nevertheless, the underlying potential of these biomaterials to reverse osteoporosis needs further research.

Nanofiber Scaffold Based on Polylactic Acid-Polycaprolactone for Anterior Cruciate Ligament Injury.

Anterior Cruciate Ligament (ACL) injuries are becoming more prevalent in athletes. Anterior Cruciatum Ligament Reconstruction (ACLR) surgery was used to treat ACL injuries and resulted in a recurrence rate of 94% due to the biomechanically repaired tissue being weaker than the original tissue. As a result, biodegradable artificial ligaments must be developed that can withstand mechanical stress during neoligament formation and stabilize the ACL. The purpose of this study is to determine the effect of composition variations in polylactic acid (PLA) and polycaprolactone (PCL) used as ACL nanofiber scaffolds on ultimate tensile strength (UTS) and modulus of elasticity, fiber diameter, cytotoxicity level, and degradation level, as well as the PLA-PCL concentration that provides the best value as an ACL scaffold. Electrospinning was used to fabricate the nanofiber scaffold with the following PLA-PCL compositions: A (100:0), B (85:15), C (80:20), D (70:30), and E (0:100) (wt%). The functional group test revealed no new peaks in any of the samples, and the ester group could be identified in the C-O bond at wave numbers 1300-1100 cm-1 and in the C=O bond at wave numbers 1750-1730 cm-1. The average fiber diameter, as determined by SEM morphology, is between 1000 and 2000 nm. The unbraided sample had a UTS range of 1.578-4.387 MPa and an elastic modulus range of 8.351-141.901 MPa, respectively, whereas the braided sample had a range of 0.879-1.863 MPa and 2.739-4.746 MPa. The higher the PCL composition, the lower the percentage of viable cells and the faster the sample degrades. All samples had a cell viability percentage greater than 60%, and samples C, D, and E had a complete degradation period greater than six months. The ideal scaffold, Sample C, was composed of PLA-PCL 80:20 (wt%), had an average fiber diameter of 827 ± 271 nm, a living cell percentage of 97.416 ± 5.079, and a degradation time of approximately 219 days.

Synthesis and Characterization of Nanohydroxyapatite-Gelatin Composite with Streptomycin as Antituberculosis Injectable Bone Substitute.

The most effective treatment for spinal tuberculosis was by eliminating the tuberculosis bacteria and replacing the infected bone with the bone graft to induce the healing process. This study aims to synthesize and characterize nanohydroxyapatite-gelatin-based injectable bone substitute (IBS) with addition of streptomycin. The IBS was synthesized by mixing nanohydroxyapatite and 20 w/v% gelatin with ratio of 40:60, 45:55, 50:50, 55:45, 60:40, 65:35, 70:30, and 75:25 ratio and streptomycin addition as antibiotic agent. The mixture was added by hydroxypropyl methylcellulose as suspending agent. FTIR test showed that there was a chemical reaction occurring in the mixture, between the gelatin and streptomycin. The result of injectability test showed that the highest injectability of the IBS sample was 98.64% with the setting time between 30 minutes and four hours after injection on the HA scaffold that represents the bone cavity and coat the pore scaffold. The cytotoxicity test result showed that the IBS samples were nontoxic towards BHK-21 fibroblast cells and human hepatocyte cells since the viability cell was more than 50% with significant difference (p-value<0.05). The acidity of the IBS was stable and it was sensitive towards Staphylococcus aureus with significantly difference (p-value<0.05). The streptomycin release test showed that the streptomycin could be released from the IBS-injected bone scaffold with release of 2.5% after 4 hours. All the results mentioned showed that IBS was suitable as a candidate to be used in spinal tuberculosis case.

Synthesis of Nanohydroxyapatite from Cuttlefish Bone (Sepia sp.) Using Milling Method.

The synthesis of nanohydroxyapatite from cuttlefish bone (Sepia sp.) has been done by using High Energy Milling (HEM) and its characterization in vitro as bone repair. This study aimed to determine the effect of the milling process on microscopic properties and mechanical properties of nano-HA through XRD, TEM, and compressive strength tests. The hydroxyapatite (HA) used in this study consisted of 1M CaCO3 from aragonite which was extracted from cuttlefish lamella bone (Sepia sp.) and 0.6 M NH4H2PO4, which was hydrothermally processed at 200°C for 12 h and then sintered at 900°C for 1h. Parameter milling includes the variation of milling time, i.e., 3 h, 6 h, and 9 h at rotational speed of 350 rpm. An increase in milling time causes a decrease in HA particle size. This is shown from the results of TEM at the milling time of 9 h with the smallest size up to 65 nm. The result of cell viability test showed that all samples are not toxic with cell viability value of >80%. The milling time of 9 h was an optimum condition with a compressive strength of 4.35952 MPa that can be applied to cancellous bone.