J Periodontal Implant Sci.  2019 Jun;49(3):148-157. 10.5051/jpis.2019.49.3.148.

Evaluation of the periodontal and pulpal healing of replanted rat molars with doxycycline root conditioning

• Nam OH
• Cheon K
• Kim MS
• Lee HS
• Choi SC

• KMID: 2454085
• DOI: http://doi.org/10.5051/jpis.2019.49.3.148

Abstract

PURPOSE
This study aimed to evaluate periodontal and pulpal healing in replanted rat teeth, preserved under different storage conditions, with or without root conditioning using doxycycline.
METHODS
A total of 40 maxillary first molars extracted from 20 Sprague-Dawley rats were stored for different durations under different conditions (5 minutes in dry storage and 60 minutes in Hank's balanced salt solution [HBSS]) and subsequently replanted. The rats were divided into 2 groups based on the use of root surface treatment: the doxycycline group (root surface treated with doxycycline) and the control group (no doxycycline treatment). Eight weeks after replantation, the animals were sacrificed, and the teeth were evaluated using micro-computed tomography (micro-CT) and histomorphometric analysis.
RESULTS
In the micro-CT analysis, the doxycycline group showed the same rate of occurrence of periapical radiolucency as was observed in the control group, but a lower degree of root resorption in teeth replanted after 60 minutes of storage in HBSS (P<0.05). In the histomorphometric analysis, the doxycycline group exhibited no improvement in either pulpal or periodontal healing of the replanted tooth after 5 minutes of dry storage, but showed a lower grade of surface root resorption (1.37±0.77) and inflammatory resorption in the teeth stored for 60 minutes in HBSS (1.33±0.71).
CONCLUSIONS
In conclusion, doxycycline improved the periodontal healing of replanted teeth stored for 60 minutes in HBSS, whereas doxycycline did not improve periodontal healing of replanted tooth after 5 minutes of dry storage. Within the limits of this study, doxycycline showed more favorable periodontal healing despite delayed replantation.

Keyword

Doxycycline; Regeneration; Replantation; Tooth injuries

MeSH Terms

Animals
Doxycycline*
Molar*
Rats*
Rats, Sprague-Dawley
Regeneration
Replantation
Root Resorption
Tooth
Tooth Injuries
Doxycycline

Figure

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  Figure 1 Flowchart of the experimental procedure.

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  Figure 2 Sections of teeth replanted after 5 minutes of dry storage. (A) Gross histological characteristics of a representative root from group I reveals newly formed PDL-like tissues attached to most of the root surface, while some areas are without PDL-like tissue attachment, showing inflammatory resorption (asterisk). (B) Sagittal micro-CT image of the root in group I shows a visible CM in the canal space. (C) High magnification of the square region of the pulp in the control group shows disruption of the odontoblastic layer (arrows). (D) High magnification of the square region of the pulp proper in the control group shows inflammatory cells and fibroblasts in the pulp proper. (E) Gross histological characteristics of a representative root from group II reveals newly formed PDL-like tissues attached to most of the root surface, while some areas fail to exhibit PDL-like tissue attachment. (F) Sagittal micro-CT image of a root from group II. (G) High magnification of the square region of the pulp in the doxycycline group shows disruption of the odontoblastic layer (arrows). (H) High magnification of the square region of the pulp in the doxycycline group shows congestion of the blood vessels in the pulp proper. (I) Histologic grade of pulpal inflammation. (J) Histologic grade of surface root resorption. (K) Histologic grade of inflammatory resorption. PDL: periodontal ligament, CT: computed tomography, CM: calcified mass.

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  Figure 3 Sections of teeth replanted after 60 minutes of storage in HBSS. (A) Gross histological characteristics of a representative root from group III reveals root resorption in almost half of the root surface area and a CM densely embedded in the pulp canal. (B) Sagittal micro-CT image of the root from group II shows severe root resorption. (C) High magnification of the square region of the root in the control group shows a moderate degree of inflammatory resorption. (D) High magnification of the square region of the root from the control group shows severe inflammatory resorption extending beyond the cementum. (E) Gross histological characteristics of a representative root from group IV shows some degree of root resorption and a CM embedded in the pulp canal. (F) Sagittal micro-CT image of a root from Group IV. (G) High magnification of the square region of the root in the doxycycline group shows severe inflammatory resorption extending beyond the cementum. (H) High magnification of the square region of the root in the doxycycline group shows PDL-like tissue attachment in the presence of surface root resorption. (I) Histologic grade of pulpal inflammation. (J) A significantly lower histologic grade of surface root resorption was achieved in the doxycycline group (P=0.009). (K) A significantly lower histologic grade of inflammatory resorption was achieved in the doxycycline group (P=0.007). HBSS: Hank's balanced saline solution, CM: calcified mass, CT: computed tomography, PDL: periodontal ligament. a)Statistically significant difference between the control and doxycycline groups (P<0.05, Mann-Whitney test).

Reference

1. Lee EU, Lim HC, Lee JS, Jung UW, Kim US, Lee SJ, et al. Delayed intentional replantation of periodontally hopeless teeth: a retrospective study. J Periodontal Implant Sci. 2014; 44:13–19.

2. Andreasen JO, Andreasen FM, Andersson L. Textbook and color atlas of traumatic injuries to the teeth. 4th ed. Copenhagen: Blackwell Munksgaard;2013.

3. Cho SY, Cheng AC. Replantation of an avulsed incisor after prolonged dry storage: a case report. J Can Dent Assoc. 2002; 68:297–300.

4. Fuss Z, Tsesis I, Lin S. Root resorption--diagnosis, classification and treatment choices based on stimulation factors. Dent Traumatol. 2003; 19:175–182.

5. McIntyre JD, Lee JY, Trope M, Vann WF Jr. Management of avulsed permanent incisors: a comprehensive update. Pediatr Dent. 2007; 29:56–63.

6. Andreasen JO. Effect of extra-alveolar period and storage media upon periodontal and pulpal healing after replantation of mature permanent incisors in monkeys. Int J Oral Surg. 1981; 10:43–53.

7. Ahn HJ, Nam OH, Lee HS, Kim EC, Cohenca N, Choi SC. Expression of inflammatory cytokines and MMPs on replanted teeth at different extra-alveolar time: an ex vivo and in vivo study. Int J Paediatr Dent. 2016; 26:301–309.

8. Andersson L, Andreasen JO, Day P, Heithersay G, Trope M, Diangelis AJ, et al. International Association of Dental Traumatology guidelines for the management of traumatic dental injuries: 2. Avulsion of permanent teeth. Dent Traumatol. 2012; 28:88–96.

9. Cvek M, Cleaton-Jones P, Austin J, Lownie J, Kling M, Fatti P. Effect of topical application of doxycycline on pulp revascularization and periodontal healing in reimplanted monkey incisors. Endod Dent Traumatol. 1990; 6:170–176.

10. Ritter AL, Ritter AV, Murrah V, Sigurdsson A, Trope M. Pulp revascularization of replanted immature dog teeth after treatment with minocycline and doxycycline assessed by laser Doppler flowmetry, radiography, and histology. Dent Traumatol. 2004; 20:75–84.

11. Yanpiset K, Trope M. Pulp revascularization of replanted immature dog teeth after different treatment methods. Endod Dent Traumatol. 2000; 16:211–217.

12. Torabinejad M, Shabahang S, Aprecio RM, Kettering JD. The antimicrobial effect of MTAD: an in vitro investigation. J Endod. 2003; 29:400–403.

13. Mohammadi Z, Abbott PV. On the local applications of antibiotics and antibiotic-based agents in endodontics and dental traumatology. Int Endod J. 2009; 42:555–567.

14. Jabarifar E, Khalighinejad N, Khademi AA, Razavi SM, Birjandi N, Badrian H, et al. Histologic evaluation of apical pulp of immature apex following extraction, surface treatment, and replantation in different storage media in dogs. Dent Traumatol. 2015; 31:118–124.

15. Cho MI, Garant PR. The effect of beta-aminoproprionitrile on the periodontal ligament: II. Radioautographic study of collagen secretion from fibroblasts. Anat Rec. 1984; 209:41–52.

16. de Carvalho FB, Andrade AS, Barbosa AF, Aguiar MC, Cangussu MC, Pinheiro AL, et al. Evaluation of laser phototherapy (λ 780 nm) after dental replantation in rats. Dent Traumatol. 2016; 32:488–494.

17. Andreasen JO, Borum MK, Jacobsen HL, Andreasen FM. Replantation of 400 avulsed permanent incisors. 4. Factors related to periodontal ligament healing. Endod Dent Traumatol. 1995; 11:76–89.

18. Pohl Y, Filippi A, Kirschner H. Results after replantation of avulsed permanent teeth. II. Periodontal healing and the role of physiologic storage and antiresorptive-regenerative therapy. Dent Traumatol. 2005; 21:93–101.

19. Preshaw PM, Hefti AF, Jepsen S, Etienne D, Walker C, Bradshaw MH. Subantimicrobial dose doxycycline as adjunctive treatment for periodontitis. A review. J Clin Periodontol. 2004; 31:697–707.

20. Ramamurthy NS, Golub LM. Diabetes increases collagenase activity in extracts of rat gingiva and skin. J Periodontal Res. 1983; 18:23–30.

21. Franco GC, Kajiya M, Nakanishi T, Ohta K, Rosalen PL, Groppo FC, et al. Inhibition of matrix metalloproteinase-9 activity by doxycycline ameliorates RANK ligand-induced osteoclast differentiation in vitro and in vivo . Exp Cell Res. 2011; 317:1454–1464.