A histological evaluation of cellular response on bone regeneration scaffold

https://doi.org/10.22146/majkedgiind.77449

Anne Handrini Dewi(1*), Dyah Listyarifah(2), Retno Ardhani(3), John Jansen(4)

(1) Department of Dental Biomedical Sciences, Faculty of Dentistry, Universitas Gadjah Mada, Yogyakarta
(2) Department of Dental Biomedical Sciences, Faculty of Dentistry, Universitas Gadjah Mada, Yogyakarta
(3) Department of Dental Biomedical Sciences, Faculty of Dentistry, Universitas Gadjah Mada, Yogyakarta
(4) Department of Biomaterials, Radboud University Nijmegen Medical Center, 6500 HB Nijmegen
(*) Corresponding Author

Abstract


Biocompatibility testing for a new implant material is necessary before clinical use. The materials for bone scaffolding must also go through biocompatibility tests both in vitro and in vivo. The purpose of this study was to compare different staining methods in histological preparations to assess the biocompatibility of the material to tissues. Twenty Wistar rats (n= 20), aged 4 months, males, weighing about 250-350 g were divided into 2 research groups, namely sub-cutaneous and bone implantation for 3, 14, and 30 days. The sample was a biodegradable material with gypsum and calcium carbonate as the basic ingredients. Histological stains were prepared in several ways using Hematoxylin Eosin (HE), Mallory, Toluidine Blue, Tartrate-resistant acid phosphatase polyclonal antibody (TRAP), and Chromogen 3,3 diaminobenzidine (DAB) after pre-performing a decalcification process with ethylenedinitril-o-tetra-acetic acid (EDTA). The observation was done using magnification ranging from 10X10 to 100X10 with a light microscope (Nikon eclipse E600) and OptiLab viewer. The results were displayed in the form of descriptions of images generated from different staining methods. Staining by the HE method is most often chosen because it is cheap and easy but the other staining methods such as Toluidine Blue, Mallory, and TRAP show better contrast. Tissue staining techniques with immunohistochemistry (DAB, VEGF) are used to evaluate cell and tissue responses by identifying specific proteins present.

Keywords


DAB; HE; mallory; scaffolding; toluidine blue; TRAP

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References

1. Yamada M, Egusa H. Review: Current bone substitutes for implant dentistry. J Prosthodont Res. 2018; 62(2):152-161. doi: 10.1016/j.jpor.2017.08.010

2. Koh TJ, DiPietro LA. Inflammation and wound healing: the role of the macrophage. Expert
Rev Mol Med. 2013; 13: e23. doi: 10.1017/S1462399411001943

3. Li Y, Chen S, Li L, Qin L, Wang X, Lai Y. Review article: Bone defect animal models for testing
efficacy of bone substitute biomaterials. J Orthop Translat. 2015; 3(3): 95-104.
doi: 10.1016/j.jot.2015.05.002

4. Listyarifah D, Susilowati R, Ana ID. Histocompatibility evaluation of combination
of gypsum with carbonated hydroxyapatite as bone substitutes in subcutaneous tissue. J
Med Sci. 2011; 43(2): 64-70.

5. Bracey DN, Jinnah AH, Willey JS, Seyler TM, Hutchinson ID, Whitlock PW, Smith TL, Danelson KA, Emory CL, Kerr BA. Investigating the osteoinductive potential of a decellularized xenograft bone substitute. Cells Tissues Organs. 2019; 207(2): 97–113. doi: 10.1159/000503280

6. Dewi AH, Ana ID, Wolke J, Jansen J. Behavior of POP-calcium carbonate hydrogrl as bone
substitute with controlled release capability: A study in rat. J biomed Mater Res Part A. 2015;
1032A : 3273-3283. doi: 10.1002/jbm.a.35460

7. Dewi AH, Ana ID, Wolke J, Jansen J. Behavior of plaster of paris-calcium carbonate
composite as bone substitute. A. study in rats, J Biomed Mater Res Part A 2013:101A:2143-
2150. doi: 10.1002/jbm.a.34513

8. Matsushige T, Kuwamoto S, Matsushita M, Wardhani LO, Yasushi Horie Y, Hayashi K,
Onago KY. RT-PCR, reverse transcription polymerase chain reaction detection of diseasespecific
fusion genes of soft tissue tumors using formalin-fixed paraffin-embedded tissues; its diagnostic usefulness and factors affecting the detection rate. acta med. 2019; 62: 115–123.
doi: 10.33160/yam.2019.03.016

9. Pröhl A, Batinic M, Alkildani S, Hahn M, Radenkovic M, Najman S, Jung O, Barbeck
M, Fernández-García M. In vivo analysis of the biocompatibility and bone healing capacity
of a novel bone grafting material combined with hyaluronic acid. Int J Mol Sci. 2021;
22(9): 4818. doi: 10.3390/ijms22094818

10. Dewi AH, Yulianto DK, Siswomihardjo W, Rochmadi R, Ana ID. Effect of dehydrothermal
treatment on the mechanical properties and biocompatibility of plaster of paris–CaCO3
hydrogel loaded with cinnamaldehyde for biomedical purposes. Nat Prod Commun. 2021;
16(1): 1–13. doi: 10.1177/1934578x20984609

11. María I, Echazú E, Sandra J, Renou, Gisela S, Alvarez, Martín F, Desimone, Olmedo DG.
Synthesis and evaluation of a chitosan–silicabased bone substitute for tissue engineering.
J Mol Sci. 2022; 23: 13379. doi: 10.3390/ijms232113379

12. Sridharan G, Akhil A Shankar AA. Review article: toluidine blue: a review of its chemistry
and clinical utility. J Oral Maxillofac Pathol. 2012; 16(2): 251-255. doi: 10.4103/0973-029X.99081

13. Listyarifah D, Al-Samadi A, Salem A, Syaify A, Salo T, Tervahartiala T, Grenier D, Nordström
D, Sorsa T, Ainola M. Infection and apoptosis associated with inflammation in periodontitis:
An immunohistologic study. Oral Dis. 2017; 23: 1144–1154. doi: 10.1111/odi.1271

14. Listyarifah D, Nieminen MT, Mäkinen LK, Haglund C, Grenier D, Häyry V, Nordström D,
Hernandez M, Yucel-Lindberg T, Tervahartiala T, Ainola M, Sorsa T, Hagström J. Treponema
denticola chymotrypsin-like proteinase is present in early-stage mobile tongue squamous cell carcinoma and related to the clinicopathological features. J Oral Pathol Med.
2018; 47(8): 764-772. doi:10.1111/jop.12729

15. Nieminen MT, Listyarifah D, Hagström J, Haglund C, Grenier D, Nordström D, Uitto V-J,
Hernandez M, Yucel-Lindberg T, Tervahartiala T, Ainola M, Sorsa T. Treponema denticola
chymotrypsin-like proteinase may contribute to orodigestive carcinogenesis through
immunomodulation. Br J Cancer. 2018; 118: 428–434. doi:10.1038/bjc.2017.409



DOI: https://doi.org/10.22146/majkedgiind.77449

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