Mimicked Periosteum Layer Based on Deposited Particle Silk Fibroin Membrane for Osteogenesis and Guided Bone Regeneration in Alveolar Cleft Surgery: Formation and in Vitro Testing

Abstract

An alveolar cleft is a critical tissue defect often treated with surgery. In this research, the mimicked periosteum layer based on deposited silk fibroin membrane was fabricated for guided bone regeneration in alveolar cleft surgery. The deposited silk fibroin particle membranes were fabricated by spray-drying with different concentrations of silk fibroin (v/v): 0.5% silk fibroin (0.5% SFM), 1% silk fibroin (1% SFM), 2% silk fibroin (2% SFM), and 1% silk fibroin film (1% SFF) as the control. The membranes were then characterized and the molecular organization, structure, and morphology were observed with FT-IR, DSC, and SEM. Their physical properties, mechanical properties, swelling, and degradation were tested. The membranes were cultured with osteoblast cells and their biological performance, cell viability and proliferation, total protein, ALP activity, and calcium deposition were evaluated. The results demonstrated that the membranes showed molecular transformation of random coils to beta sheets and stable structures. The membranes had a porous layer. Furthermore, they had more stress and strain, swelling, and degradation than the film. They had more unique cell viability and proliferation, total protein, ALP activity, calcium deposition than the film. The results of the study indicated that 1% SFM is promising for guided bone regeneration for alveolar cleft surgery.

Keywords: Silk fibroin particle; alveolar cleft; guided bone regeneration; mimicking; periosteum; spray drying.

Figure 1.

Physical appearances of the silk fibroin film (SFF) and silk fibroin membranes (SFMs): 1% SFF (a), 0.5% SFM(b), 1% SFM (c), and 2% SFM (d).

Figure 2.

FTIR spectrums of the 1% SFF and the 2%, 1%, and 0.5% SFMs.

Figure 3.

DSC thermograms of the 1% SFF and the 2%, 1%, and 0.5% SFMs.

Figure 4.

Morphologies of the silk fibroin film and silk fibroin membranes: 1% SFF (a), 0.5% SFM (b), 1% SFM (c), and 2% SFM (d).

Figure 5.

Pore size distributions of the 0.5% SFM, 1% SFM, and 2% SFM: large pore size distribution (a) and small pore size distribution (b).

Figure 6.

Percentages of swelling of the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM.

Figure 7.

Degradation of the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM after days 1, 3, 5, 7, 14, and 21. * p < .05.

Figure 8.

Mechanical properties of the membranes in both the wet and dry states: stress at maximum load (a), strain at maximum load (b), and Young’s modulus (c). * p < .05.

Figure 9.

Cell viability on the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM at days 3, 5, and 7. Scale bar: 100 µm.

Figure 10.

Cell proliferation of the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM at days 1, 3, 5, and 7. * p < .05.

Figure 11.

Total protein content of the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM after cell curing at days 7, 14, and 21. * p < .05.

Figure 12.

ALP activity of 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM at days 7, 14, and 21. * p < .05.

Figure 13.

Calcium deposition on the 1% SFF, 0.5% SFM, 1% SFM, and 2% SFM. Scale bar: 100 µm.

Figure 14.

Structure of alveolar cleft (a), alveolar cleft surgery with membrane (b).