THE EFFECTIVITY OF SCAFFOLD FROM ANADARA GRANOSA SHELL-STICHOPUS HERMANNI ON BLOOD VESSEL COUNTS AFTER TOOTH EXTRACTION

Background: Alveolar bone resorption can occur after tooth extraction. One source of bone scaffold material is hydroxyapatite and tricalcium phosphate (HA-TCP). The shell of Anadara granosa can be synthesized to be HA-TCP. Hyaluronic acid, which is widely contained in Stichopus hermanni, can stimulate endothelial progenitor cells for the healing process. Purpose: This research aims to prove the Effectivity of scaffold from Anadara granosa shell-Stichopus hermanni on blood vessel counts after tooth extraction. Methods: The sample in this study was male Wistar rats divided into four study groups randomly. Each group consists of 6 rats. Extraction was carried out on the mandibular first incisor, then was given gelatin as a placebo in the control group, treatment groups given scaffold from Anadara granosa shells (AG), and scaffold combination from Anadara granosa shells-Stichopus hermanni with concentrations of 0.4%-0.8%. (AGSH1-AGSH2). Development of HA-TCP synthesized from Anadara granosa combined with Stichopus hermanni extract for biomedical scaffolds using the freeze-dried method. Observations were made three days after tooth extraction. Data analysis has used one-way ANOVA followed by Tukey HSD (p<0.05). Results: The results of the ANOVA test followed by the HSD-Tukey test showed a significant difference between C (7.33±1.51) and AGSH1 (11.50±1.38), AGSH2 (21.17±1.94), AG (9.33±1.63) and AGSH1 with AGSH2, but between AG and, AGSH1 no significant differences. Conclusions: Combination scaffold from Anadara granosa shell and Stichopus hermanni 0.8% effectively increased blood vessel counts after tooth extraction. INTRODUCTION Indonesian Basic Health Research 2013 showed index M-T (missing teeth) scores 2.9 which expresses the number of permanent teeth extraction is 290 for 100 patients of tooth decay. This number considered high as if we compare it to F-T (filling teeth) index which scores 0.08.1 Posttooth extraction, in general, are likely to cause the decreasing of alveolar ridge height and morphology changes, which in turn, can lead into bone resorption and moreover will be disruption in the stomatognathic system.2 Post-tooth extraction, the socket healing process involves several phases, namely the inflammatory phase, the proliferative phase, and the remodeling phase. The proliferation phase will begin on the third day after tissue injury. This phase was characterized by fibroblasts migration, * Department of Oral Biology, Faculty of Dentistry, Universitas Hang Tuah ** Department of Prosthodontic, Faculty of Dentistry, Universitas Hang Tuah *** Department of Oral Pathology and Maxilofacial, Faculty of Dental Medicine, Airlangga University ****Department of Pharmacology, Faculty of Veterinary Medicine, Airlangga University Correspondence: rima.parwatisari@hangtuah.ac.id


Indonesian Basic Health Research 2013
showed index M-T (missing teeth) scores 2.9 which expresses the number of permanent teeth extraction is 290 for 100 patients of tooth decay.
This number considered high as if we compare it to F-T (filling teeth) index which scores 0.08. 1 Posttooth extraction, in general, are likely to cause the decreasing of alveolar ridge height and morphology changes, which in turn, can lead into bone resorption and moreover will be disruption in the stomatognathic system. 2 Post-tooth extraction, the socket healing process involves several phases, namely the inflammatory phase, the proliferative phase, and the remodeling phase. The proliferation phase will begin on the third day after tissue injury. This phase was characterized by fibroblasts migration, Sari/ Kusumawardani/ Damaiyanti/ Rahayu/ Sudjarwo 12 ODONTO Dental Journal. Volume 8. Nomor 2. Desember 2021 extracellular matrix deposition, collagen synthesis, angiogenesis, and granulation tissue formation. 3 Angiogenesis plays a crucial role in the wound healing process by supplying oxygen and nutrition to support cell metabolism. 4,5 Angiogenesis is directly involved in maintaining functions of impacted tissue and organ. 6  Wistar rats were acclimatized for one week and rats were fasted overnight before conducting the study. Randomization of samples into groups was carried out before weighing and marking.
Furthermore, the rat was anesthetized using 10% ketamine at a dose of 0.1 cc/kg body weight and xylazine 0.01 cc per 100 g body weight in the right upper thigh (intramuscular). 19 Make sure the rat has been affected by anesthesia marked by a corneal reflex missing before the reflexes of the foot Multiple comparison results HSD showed the significant differences of C control group to AGSH1-2 group , AG group and AGSH1 group to AGSH2 group. Insignificant differences showed between C group to AG group and AG group to AGSH1 group (Table 1). Vascular observation showed in group K = 7.33 ± 1.51; AG = 9.33 ± 1.63; AGSH1=11.50±1.38; AGSH2=21.17±1.94 ( Figure   2).

Figure 1. Histologic section of blood vessels at the apical 2/3 socket
A. K group (Control group given scaffold from gelatin); B. AG group (group given scaffold from Anadara granosa shell); C. AGSH1 group (group given scaffold from Anadara granosa shell-Stichopus hermanni 0,4% combination); D. AGSH2 Group (The group given scaffold 0.8% Anadara granosa-Stichopus hermanni 0.8% combination scaffold) Multiple comparison results HSD showed the significant differences of C control group to AGSH1-2 group , AG group and AGSH1 group to AGSH2 group. Insignificant differences showed between C group to AG group and AG group to AGSH1 group (Table 1).  Note: This table shows the comparison of the mean number of blood vessels between groups. The average number of blood vessels in group AGSH greater than groups K and AG. The greatest of blood vessels is AGSH2. These can be seen at control group, where blood vessels still can be found ( Figure 1A).

DISCUSSION
HA-TCP application from the synthesized of Anadara granosa shell in this research, functionate as a scaffold. Scaffold from HA-TCP expected to have mineral cavities that is connected one to another, thus allows the adhesion of mesenchyme cell to differentiate forming a new blood vessels. 23 In this study, we can conclude that the increased blood vessels was found in AG group, although in the multiple comparison HSD test, there was no significant differences with C group. This means that scaffold from HA-TCP alone was not enough to improve the blood vessels count significantly to accelerate angiogenesis process.