Cellulose Acetate Nanofibers: Incorporating Hydroxyapatite (HA), HA/Berberine or HA/Moghat Composites, as Scaffolds to Enhance In Vitro Osteoporotic Bone Regeneration.

Nadia Z Shaban,Marwa Y Kenawy,Doaa A Ghareeb,Mona M Abd El-Latif,Nahla A Taha

doi:10.3390/polym13234140

Nadia Z Shaban, Marwa Y Kenawy + Show 3 more

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https://doi.org/10.3390/polym13234140

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Abstract

The specific objective of this study was to stabilize a simple valid method to prepare pure nanorod hydroxyapatite (HA) mixed with berberine chloride (BER) and Moghat water extract (ME) as composites for incorporation into cellulose acetate (CA) nanofibers to be used as novel bone scaffolds and to determine their efficacy in bone regeneration process In Vitro. Preparation of HA/BER and HA/ME composites were performed by mixing powders using the ball-milling machine. The HA, HA/BER, and HA/ME composites at a concentration of 6.25, 12.5, 25, 50, 100, and 200 mg were mixed with CA solution (13%), then the fiber was formed using electrospinning technique. The properties of the obtained CA fibers were investigated (SEM, TEM, EDX, FTIR, TGA, water uptake, porosity, and mechanical tests). The efficacy of HA and HA composites loaded into CA nanofiber on osteoblast and osteoclast differentiation were measured by tacking ALP, osteocalcin, TRAcP, calcium, and total protein concentration. Moreover, their effects on cell differentiation (CD90 and PARP- ɣ) and death markers (GSK3b, MAPK, Wnt-5 and β-catenin) were evaluated by using ELISA and qPCR. The obtained TEM results indicated that the continuous CA and CA/HA composites electrospun fibers have ultrafine fiber diameters of about 200 nm and uniform distribution of discrete n-HA clusters throughout. In addition, hydrocortisone (HCT) was found to increase the formation of adipocytes and osteoclastic markers CD90 and p38-MAPK which indicated the bone lose process take placed. Treatment with CA loaded with HA, HA/BER or HA/ME decreased CD90, Wnt-5, PARP- ɣ, GSK3b and p38-MAPK associated elevation of osteogenic markers: ALP and osteocalcin. Moreover, HCT overexpressed RANKL and down expressed Osterix gene. Treatment with CA/HA/BER or CA/HA/ME downregulated RANKL and upregulated Osterix associated with a reduction in RANKL/OPG ratio, at p < 0.05. In conclusion, novel CA composite nanofibers (CA/HA/BER and CA/HA/ME) reversed the HCT adverse effect on osteoblast cell death through canonical and non-canonical pathways regulated by Wnt/β-catenin and Wnt/Ca(2+) pathways. Furthermore, our data confirmed that the novel scaffolds create a crosstalk between RUNX-2, RANKL, p38-MAPK, and Wnt signals which positively impact bone regeneration process. Treatment with CA/HA/BER is better compared to the treatment with CA/HA/ME. Nevertheless, both are considered as alternative biomaterial scaffolds with a potential for biomedical applications in the field of bone tissue engineering.

Highlights

Tissue engineering is a vital area of regenerative medicine, with rapidly expanding research directed at preparing recent biomaterial matrices with adjusted properties to get functional tissues for specific applications [1]
Cross-sectional round fibers were obtained and there was no presence of any kind of HA or HA composites aggregates on the surface of these fibers, implying that the as-loaded HA, HA/berberine chloride (BER), and HA/Moghat water extract (ME) were perfectly incorporated well within the CA13 fibers
While for CA13/HA50, CA13/HA/BER100, and CA13/HA/ME100, (e) roughness and irregularities are observed, due to(f) surface the presence of the HA, HA/BER, and HA/ME composites loaded on the surface and inside the nanofiber mats (Figure 1)

Summary

Introduction

Tissue engineering is a vital area of regenerative medicine, with rapidly expanding research directed at preparing recent biomaterial matrices with adjusted properties to get functional tissues for specific applications [1]. Designing a biocompatible scaffold with optimal characteristics associated with the above is a major key element of successful tissue engineering strategies [2]. These developed scaffolds should have satisfactory biocompatibility and structure similar to that of the native extracellular matrix (ECM) in order to enable cells to grow and differentiate into specific tissues similar to their natural counterpart [3]. Bone healing is regeneration defined as a complex and wellorchestrated process of biological events of bone induction and conduction to optimize skeletal repair and re-store skeletal function [5]. To stimulate or augment bone regeneration of large bone defects, bone grafting such as autologous bone grafts and allografts is a commonly performed surgical procedure [6]

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