Comprehensive In Vitro Testing of Calcium Phosphate-Based Bioceramics with Orthopedic and Dentistry Applications

Abstract

Recently, a large spectrum of biomaterials emerged, with emphasis on various pure, blended, or doped calcium phosphates (CaPs). Although basic cytocompatibility testing protocols are referred by International Organization for Standardization (ISO) 10993 (parts 1–22), rigorous in vitro testing using cutting-edge technologies should be carried out in order to fully understand the behavior of various biomaterials (whether in bulk or low-dimensional object form) and to better gauge their outcome when implanted. In this review, current molecular techniques are assessed for the in-depth characterization of angiogenic potential, osteogenic capability, and the modulation of oxidative stress and inflammation properties of CaPs and their cation- and/or anion-substituted derivatives. Using such techniques, mechanisms of action of these compounds can be deciphered, highlighting the signaling pathway activation, cross-talk, and modulation by microRNA expression, which in turn can safely pave the road toward a better filtering of the truly functional, application-ready innovative therapeutic bioceramic-based solutions.

Potential Application of Protamine for Antimicrobial Biomaterials in Bone Tissue Engineering

“…Generally, antibacterial ability conflicts with biocompatibility. Therefore, to evaluate the biocompatibility of protamine/HAp, in vitro cytotoxicity tests were carried out according to ISO 10993-5:2009 “Tests for Cytotoxicity—In vitro Methods” [ 20 ]. Human osteoblastic cells (MG-63) were cultured in extracts from various kinds of powders (0.1 mg·mL −1 ) for 24 and 72 h. As shown in Figure 7 , the results of the MTT assay at 24 and 72 h showed that no significant changes of cell viability were seen in concentrations ranging from 125 to 500 μg·mL −1 .…”

“…To evaluate the antibacterial activity, Escherichia coli (E. coli, ATCC 27325) and Staphylococcus aureus (S. aureus, ATCC 6538) were used in this study. Bacteria were grown in a culture broth (nutrient USA) supplemented with 10% heat-inactivated fetal bovine serum (Sigma, St. Louis, MO, USA) and 0.1% antibiotics (penicillin-streptomycin solution, Sigma) in a humidified atmosphere containing 5% CO 2 at 37 • C. According to ISO 10993-5:2009 "Tests for Cytotoxicity-In vitro Methods" [20], extracts were obtained in a separate culture medium (0.1 g•mL −1 of the culture medium for 24 h at 37 • C). For cytotoxicity test, cells were seeded at 5 × 10 3 cells•well -1 on 96-well plates and cultured for 24 h at 37 • C/5% CO 2 .…”

Is THP‐1 viability affected by the crystallinity of nanostructured carbonated hydroxyapatites?

“…Because THP‐1 cells, a cell line of human monocyte cells, were used in the present study, we could evaluate not only the behavior of cells against the biomaterials but also the behavior of cells involved in the inflammatory immune process for protecting tissues from toxins and initiating the resolution of inflammation 30 . Monocyte recruitment and differentiation may occur in the tissue in response to calcium phosphate‐based biomaterials 31 . Additionally, macrophages are the principal cells responsible for mediating the proinflammatory response toward implant debris as a precursor of osteoclasts 32 …”

“…30 Monocyte recruitment and differentiation may occur in the tissue in response to calcium phosphate-based biomaterials. 31 Additionally, macrophages are the principal cells responsible for mediating the proinflammatory response toward implant debris as a precursor of osteoclasts. 32 Previous studies showed the variable effect of the size and shape of nanostructured HA particles on cellular viability.…”

Analysis of cell-biomaterial interaction through cellular bridge formation in the interface between hGMSCs and CaP bioceramics

“…The process of bone regeneration integrated with bioceramics consists on different steps that can be summarized as a complex and multistage process resulting from a plethora of biological stages; including osteogenesis, angiogenesis and inflammatory responses. All these processes are essential for the recovery of tissue homeostasis and function 13 but for them to occur a good biocompatilibity between the bioceramic and the nearby tissue is essential 12,16 .…”