Effect of bovine hydroxyapatite composite with secretome under normoxia and hypoxia conditions on inflammatory parameters in massive bone defect of rabbit radius bone

Abstract

Hydroxyapatite as a scaffold is capable of producing good bone regeneration formation. Incorporating secretome into scaffolds optimizes the bone healing process. The increase in proinflammatory, anti-inflammatory, and growth factors is one of the key factors in bone healing. In this study, we measured the levels of IL-6, IL-10, and FGF-2 to determine the effectiveness of bovine hydroxyapatite with secretome from normoxia and hypoxia on bone healing. This animal study employed a pure experimental research design, utilizing a post-test-only control group design. Bone marrow mesenchymal stem cells from rabbit thigh bones were used to derive secretomes under hypoxic and normoxic conditions. Bovine bone-derived hydroxyapatite (BHA) was treated with secretomes under both conditions. Rabbits’ radius bones were implanted with BHA alone, BHA with normoxic secretome, and BHA with hypoxic secretome, then observed for 30 and 60 days. Levels of IL-6, IL-10, and FGF-2 were examined on days 30 and 60. On the 30th day, there was a significant increase in the levels of FGF-2, IL-6, and IL-10, with a dominance of strongly positive levels in BHA alone. However, on the 60th day, the levels of FGF-2, IL-6, and IL-10 started to decrease in all groups, with a dominance of moderately positive levels. Statistical tests showed significant results in all groups on days 30 and 60 (p < .05). Among the three groups, the best levels of growth factors and pro-inflammatory factors, and the lowest levels of anti-inflammatory factors were found in the BHA alone group on evaluation day 30.

Keywords

Massive bone defect bovine hydroxyapatite secretome normoxic hypoxic

Get full access to this article

View all access options for this article.

References

Rather

Jhala

Vasita

. Dual functional approaches for osteogenesis coupled angiogenesis in bone tissue engineering. Mater Sci Eng C 2019; 103: 109761, DOI: 10.1016/j.msec.2019.109761.

Alizadeh-Osgouei

Wen

. A comprehensive review of biodegradable synthetic polymer-ceramic composites and their manufacture for biomedical applications. Bioact Mater 2019; 4: 22–36, DOI: 10.1016/j.bioactmat.2018.11.003.

Mahyudin

Utomo

Suroto

, et al. Comparative effectiveness of bone grafting using xenograft freeze-dried cortical bovine, allograft freeze-dried cortical New Zealand white rabbit, xenograft hydroxyapatite bovine, and xenograft demineralized bone matrix bovine in bone defect of femoral diaphysis of white rabbit: experimental study in vivo. Int J Biomater 2017; 2017: 7571523–7571529.

Tajima

Tobita

Orbay

, et al. Direct and indirect effects of a combination of adipose-derived stem cells and platelet-rich plasma on bone regeneration. Tissue Eng 2015; 21(5–6): 895–905, DOI: 10.1089/ten.tea.2014.0336.

Ahmed

NE-MB

Murakami

Kaneko

, et al. The effects of hypoxia on the stemness properties of human dental pulp stem cells (DPSCs). Sci Rep 2016; 6(1): 35476, DOI: 10.1038/srep35476.

Woodbury

Ikezu

. Fibroblast growth factor-2 signaling in neurogenesis and neurodegeneration. J Neuroimmune Pharmacol 2014; 9(2): 92–101, DOI: 10.1007/s11481-013-9501-5.

Ornitz

Itoh

. The fibroblast growth factor signaling pathway. Wiley Interdiscip Rev Dev Biol 2015; 4(3): 215–266, DOI: 10.1002/wdev.176.

Rose-John

. Interleukin-6 family cytokines. Cold Spring Harbor Perspect Biol 2018; 10(2): a028415, DOI: 10.1101/cshperspect.a028415.

Scheller

Chalaris

Schmidt-Arras

, et al. The pro- and anti-inflammatory properties of the cytokine interleukin-6. Biochim Biophys Acta 2011; 1813(5): 878–888, DOI: 10.1016/j.bbamcr.2011.01.034.

10.

Saraiva

O’Garra

. The regulation of IL-10 production by immune cells. Nat Rev Immunol 2010; 10(3): 170–181, DOI: 10.1038/nri2711.

11.

King

Balaji

, et al. Regenerative wound healing: the role of interleukin-10. Adv Wound Care 2014; 3(4): 315–323, DOI: 10.1089/wound.2013.0461.

12.

Charoenlarp

Rajendran

Iseki

. Role of fibroblast growth factors in bone regeneration. Inflamm Regen 2017; 37(1): 10, DOI: 10.1186/s41232-017-0043-8.

13.

O’Keefe

Jacobs

Chu

, et al. and American Academy of Orthopaedic Surgeons. Orthopaedic basic science: foundations of clinical practice. Rosemont: American Academy of Orthopaedic Surgeons, 2013.

14.

Idzik

Poloczek

Skrzep-Poloczek

, et al. The effects of 21-day general rehabilitation after hip or knee surgical implantation on plasma levels of selected interleukins, VEGF, TNF-α, PDGF-BB, and eotaxin-1. Biomolecules 2022; 12(5): 605, DOI: 10.3390/biom12050605.

15.

Maruyama

Rhee

Utsunomiya

, et al. Modulation of the inflammatory response and bone healing. Front Endocrinol 2020; 11: 386, DOI: 10.3389/fendo.2020.00386.

16.

Kaiser

Prystaz

Vikman

, et al. Pharmacological inhibition of IL-6 trans-signaling improves compromised fracture healing after severe trauma. Naunyn-Schmiedeberg’s Arch Pharmacol 2018; 391(5): 523–536, DOI: 10.1007/s00210-018-1483-7.

17.

Yang

, et al. Interaction between mesenchymal stem cells and immune cells during bone injury repair. Int J Mol Sci 2023; 24(19): 14484, DOI: 10.3390/ijms241914484.

18.

Prystaz

Kaiser

Kovtun

, et al. Distinct effects of IL-6 classic and trans -signaling in bone fracture healing. Am J Pathol 2018; 188(2): 474–490, DOI: 10.1016/j.ajpath.2017.10.011.

19.

El-Fetiany

Kassem

El-Barbary

, et al. Evaluation of plasma basic fibroblast growth factor (bFGF) in primary knee osteoarthritis patients. The Egyptian Rheumatologist 2017; 39(1): 33–37, DOI: 10.1016/j.ejr.2016.03.006.