Tissue-Engineered Hydroxyapatite Bone Scaffold Impregnated with Osteoprogenitor Cells Promotes Bone Regeneration in Sheep Model

Abstract

BACKGROUND: Managing massive bone defects, a great challenge to orthopaedics reconstructive surgery. The problem arise is the supply of suitable bone is limited with many complications. Tissue-engineered hydroxyapatite bone (TEHB) scaffold impregnated with osteoprogenitor cells developed as an alternative to promote bone regeneration. METHODS: This animal protocol has been approved by Universiti Kebangsaan Malaysia Animal Ethical Committee. The TEHB scaffold prepared from hydroxyapatite using gel casting method. A total of six adolescent female sheep were chosen for this study. Later, all the sheep were euthanized in a proper manner and the bone harvested for biomechanical study. Bone marrow was collected from iliac crest of the sheep and bone marrow stem cells (BMSCs) isolated and cultured. BMSCs then cultured in osteogenic medium for osteoprogenitor cells development and the plasma collected was seeded with osteoprogenitor cells mixed with calcium chloride. Bone defect of 3 cm length of tibia bone created from each sheep leg and implanted with autologous and TEHB scaffold in 2 different groups of sheep. Wound site was monitored weekly until the wound completely healed and conventional X-ray performed at week 1 and 24. Shear test was conducted to determine the shear force on the autologous bone and TEHB scaffold after implantation for 24 weeks. RESULTS: All of the sheep survived without any complications during the study period and radiograph showed new bone formation. Later, the bone harvested was for biomechanical study. The highest shear force for the autologous group was 13 MPa and the lowest was 5 MPa while for the scaffold group, the highest was 10 MPa and the lowest was 3 MPa. Although, proximal and distal interface of autologous bone graft shows higher shear strength compared to the TEHB scaffold but there is no significant difference in both groups, p value [ 0.05. Histologically in both proximal and distal interface in both arms shows bone healing and woven bone formation. CONCLUSION: TEHB scaffold impregnated with osteoprogenitor cells has the potential to be developed as a bone substitute in view of its strength and capability to promote bone regeneration.

In Vivo Bone Tissue Engineering Strategies: Advances and Prospects

“…Previous studies have revealed key components of BTE [ 21 , 22 , 23 , 24 , 25 , 26 ]. According to these studies successful bone regeneration relies on the combination of (1) a bioactive scaffold [ 27 , 28 , 29 , 30 , 31 ]; (2) osteoprogenitor cells [ 21 , 26 , 32 , 33 , 34 , 35 ]; (3) growth factors (GF) [ 23 , 36 , 37 , 38 , 39 , 40 ] and (4) adequate vascularization [ 41 , 42 , 43 ] ( Figure 1 ).…”

“…Previous studies have revealed key components of BTE [21][22][23][24][25][26]. According to these studies successful bone regeneration relies on the combination of (1) a bioactive scaffold [27][28][29][30][31]; (2) osteoprogenitor cells [21,26,[32][33][34][35]; (3) growth factors (GF) [23,[36][37][38][39][40] and (4) adequate vascularization [41][42][43] (Figure 1). Natural polymers are particularly effective for regenerative medicine application for two main reasons.…”

Long-Bone-Regeneration Process in a Sheep Animal Model, Using Hydroxyapatite Ceramics Prepared by Tape-Casting Method

“…There are many problems and obstacles associated with these procedures, e.g., treatment is limited by bone volume, recurrent fractures, soreness, associated complications such as infection, local haematoma, poor integration, donor site shortage, prolonged surgery time, donor site morbidity, and vascular problems [ 3 , 4 , 5 ]. Generally, allografts have low osteogenicity, low mechanical resistance, can evoke immunologic reactions, and pose a risk of graft rejection [ 6 , 7 ]. Therefore, a more sustainable and long-term treatment plan is necessary.…”

“…HA was reported as nontoxic, only minimally inflammatory, and osteoconductive [ 31 , 32 ]. Many previous findings have shown that HA has a very good biocompatibility, biomimetic character [ 6 , 33 , 34 ], and the functional characteristics that facilitate cell growth and the formation of new bone. Among the various synthetic biomaterials, HA gives the best results [ 35 ].…”

Performance of Polydioxanone-Based Membrane in Association with 3D-Printed Bioceramic Scaffolds in Bone Regeneration

“…This fact gives HA a high mechanical resistance and more significant slowness in its bone replacement process [ 15 , 16 ]. HA nanoparticles promote the adhesion, proliferation, and differentiation of osteoprogenitor cells and are considered bioceramic with potent osteoconductive action [ 13 , 20 , 21 , 22 , 23 ].…”