Abstract
The wound healing process was analyzed using male ICR germ-free (Gf) mice and their conventionalized (Cvz) counterparts to determine whether intestinal flora has any influence on the wound healing of a host. A longitudinal skin incision of about 35 mm was made in the dorsum of each mouse, and six interrupted sutures with 5–0 monofilament nylon thread laid for wound closure. Mice from both groups were killed on the 3rd, 7th, and 14th postoperative days, and the tensile strength of the healing wound and hydroxyproline (HP) concentration in the biologically active zone were measured as parameters of wound healing. The daily nitrogen balance and urinary creatinine excretion were also measured for 8 days. The Cvz mice showed greater tensile strength of the wound on the 3rd and 7th postoperative days, and a higher ratio of HP concentration, being wound tissue/control tissue, than their counterparts. The Cvz mice also thrived more prominently than the Gf mice, showing a positive nitrogen balance with limited urinary creatinine excretion. These results suggest the enhancing effect of normal intestinal flora on wound healing, probably through the beneficial nutritional effects supplied by the bacteria.
Similar content being viewed by others
References
Nichols RL, Condon RE (1971) Preoperative preparation of the colon: A collective review. Surg Gynecol Obstet 132:323–337
Goligher JC (1980) Preoperative care. Surgery of the anus, rectum and colon, 4th edn. Bailliere Tindall London, pp 447–450
Roediger WEW (1980) Role of anaerobic bacteria in the metablic welfare of the colonic mucosa in man. Gut 21:793–798
Roediger WEW (1982) Utilization of nutrients by isolated epithelial cells of the rat colon. Gastroenterology 83:424–429
Coates ME, Hewitt D, Salter DN (1971) Protein metabolism in the germ-free and conventional chick. In: Heneghan JB (ed) Germfree research, biological effect of gnotobiotic environments. Academic, New York, pp 291–295
Wrong O (1978) Nitrogen metabolism in the gut. Am J Clin Nutr 31:1587–1593
Yamanaka M, Nomura T (1980) Relation between protein contents of diets and biological values in germ-free rats. J Nutr Sci Vitaminol 26:323–326
Tsuda M, Saheki S, Takada S, Kusumi T, Okubo T, Sase M, Katsunuma T (1981) Effect of intestinal flora on the metabolism of amino acids and proteins in the rat and mouse. In: Sasaki S, Ozawa A, Hashimoto K (eds) Recent advances in germ-free research. Tokai University Press, Tokyo, pp 237–242
Ishibashi T, Takahashi M, Kametaka M, Takenaka A, Ozaki A, Yamamoto T, Mitsuoka T (1983) Nutrition and intestinal flora of chicks (in Japanese with English abstract). In: Mitsuoka T (ed) Intestinal flora and nutrition. Japan Scientific Societies Press, Tokyo, pp 157–179
Honma C, Takahashi M, Kametaka M, Yamanaka M (1987) Amino acid pattern of gastrointestinal, contents in germ-free and conventionalized rats (in Japanese with English abstract). Nippon Eiyo Syokuryo Gakkaishi (J Jpn Soc Nutr Food Sci) 40:411–415
Takahashi M (1990) Protein metabolism (in Japanese). In: Mitsuoka T (ed) Chonaisaikingaku (Intestinal Bacteriology) Asakura Syoten, Tokyo, pp 256–262
Peterson JM, Barbul A, Breslin RJ, Wasserkrug HL, Efron G (1987) Significance of T-lymphocytes in wound healing. Surgery 102:300–305
Browder W, Williams D, Lucore P, Pretus H, Jones E, McNamee R (1988) Effect of enhanced macrophage function of early wound healing. Surgery 104:224–230
Efron JE, Frankel HL, Lazarou SA, Wasserkrug HL, Burbul A (1990) Wound healing and T-lymphocytes. J Surg Res 48:460–463
Barbul A (1990) Immune aspects of wound repair. Clin Plast Surg 17:433–442
Shimada Y, Kaji K (1992) Cytokines in wound healing (in Japanese). Rinsho Menneki (Clin Immunol) 24:145–152
Orcutt RP, Otis AP, Alliger H (1981) AlcideTM: An alternative sterilant to peracetic acid. In: Sasaki S, Ozawa A, Hashimoto K (eds) Recent advances in, germ-free research. Tokai University Press, Tokyo, pp 79–81
Mitsuoka T (1980) World of intestinal bacteria. Methods of analysis of intestinal flora (in Japanese). Sobunsya, Tokyo, pp 53–92
Crawford DT, Bains JW, Ketcham AS (1965) A standard model for tensiometric studies. J Surg Res 5:265–269
Prockop DJ, Udenfriend S (1960) A specific method for the analysis of, hydroxyproline in tissue and urine. Anal Biochem 1:228–239
Juva K, Prockop DJ (1966) Modified procedure for the assay of H3-or C14-labeled hydroxyproline. Anal Biochem 15:77–83
Konstantinides FN, Boehm KA, Radmer WJ, Storm MC, Adderly JT, Weisdorf SA, Cerra FB (1988) PyrochemiluminescenceTM: Real-time, cost-effective method for determining total urinary nitrogen in clinical nitrogen-balance studies. Clin Chem 34:2518–2520
Heinegard D, Tiderstrom G (1973) Determination of serum creatinine by a direct colorimetric method. Clin Chim Acta 43:305–310
Aliqureshi G, Van Den Berg S, Gutierrez A, Bergstrom J (1984) Determination of histidine and 3-methylhistidine in physiological fluids by high-performance liquid chromatography. J Chromatogr 297:83–89
Hansbrough J, Peterson V, Zapata-Sirvent R, Claman HN (1984) Postburn immunosuppression in an animal model. II: Restoration of cell-mediated immunity by immunomodulating drugs. Surgery 95:290–296
Miyakawa M, Isomura N, Shirasawa H, Yokoi K (1958) Wound healing in germ-free animals. Acta Path Jpn, 8:79–97
Tipton JB, Dingman RO (1966) Some aspects of wound healing in the germ-free animal. Plast Reconstr Surg 38:499–506
Donati RM, Frank DW, Stromberg LR, McLaughlin MM (1971) The effect of the germ-free state on wound healing. J Surg Res 11:163–172
Rovin S, Costich ER, Fleming JE, Gordon HA (1965) Healing of tongue wounds in germ-free and conventional mice. Arch Pathol 79:641–643
Brody GL, Bishop JE, Abrams GD (1966) Normal flora and collagen production. Arch Pathol 81:268–270
Gordon HA, Pesti L (1971) The gnotobiotic animal as a tool in the study of host microbial relationships. Bacteriological Reviews 35:390–429
Mastboom WJB, Hendriks T, de Boer HHM (1989) Collagen changes around intestinal anastomoses in germ-free rats. Br J Surg 76:797–801
Ito K (1990) Germ-free animals (in Japanese). In: Mitsuoka T (ed) Chonaisaikingaku (Intestinal Bacteriology). Asakura Syoten, Tokyo, pp. 145–149
Freter R, Abrams GD, Aranki A (1973) Patterns of interaction in gnotobiotic mice among bacteria of a synthetic “normal” intestinal flora. In: Heneghan JB (ed) Germ-free research, biological effect of gnotobiotic environments. Academic New York, pp 429–433
Levenson SM, Kan-Gruber D, Gruber C, Molnar J, Seifter E (1983) Wound healing accelerated byStaphylococcus aureus. Arch Surg 118:310–320
Brunel A, Gouet PH (1981) Kinetic bacterial implantation in the intestine of newborn rats. In: Sasaki S, Ozawa A, Hashimoto K (eds) Recent advances in germ-free research. Tokai University Press, Tokyo, pp 185–188
Ito K, Mitsuoka T, Freter R (1987) Mechanisms that control bacterial populations of mouse intestinal flora in gnotobiotic mice and in continuous flow cultures (in Japanese with English abstract). In: Mitsuoka T (ed) Methodology for research in intestinal flora. Japan Scientific Societies Press, Tokyo, pp 107–136
Dunphy JE, Udupa KN (1955) Chemical and histochemical sequences in the normal healing of wounds. New Eng J Med 253:847–851
Rollandelli RH, Koruda MJ, Settle RG, Rombeau JL (1986) Effects of intraluminal infusion of short-chain fatty acids on the healing of colonic anastomosis in the rat. Surgery 100: 198–203
Okada M, Konishi F, Kataoka T, Kashiwagi H, Saito Y, Kanazawa K, Hakamata Y, Hayashi N (1992) Intestinal flora enhances wound healing in mice (in Japanese) Saishin Igaku (Progress in Medicine) 12:2871–2873
Adamsons RJ, Musco F, Enquist IF (1966) The chemical dimensions of a healing incision. Surg Gynec Obstet 123:515–521
Maruyama K, Shigemori C, Miyoshi N, Aida S, Inayama S, Sibata T (1980) A study of biochemical active zone in the colonic anastomosis (in Japanese) Saishin Igaku (Progress in Medicine) 35:1726–1728
Hunt TK, Hawley PR, Hale J, Goodson W, Thakral KK (1980) Colon repair: The collagenous equilibrium. In: Hunt TK (ed) Wound healing and wound infection. Theory and surgical practice. Appleton-Century-Crofts. New York, pp. 153–159
Yokota H (1981) Intestinal absorption of nutrients in germ-free chicken. In: Sasaki S, Ozawa A, Hashimoto K (eds) Recent advances in germ-free research. Tokai University Press Tokyo, pp 343–346
Heneghan JB (1963) Influence of microbial flora on xylose absorption in rats and mice. Am J Physiol 205:417–420
Herskovic T, Katz J, Floch MH, Spencer RP, Spiro HM (1967) Small intestinal absorption and morphology in germ-free, monocontaminated, and conventionalized mice. Gastroenterology 52: 1136
Suzuki K, Mitsuoka T (1976) Urease activity of intestinal anaerobes (in Japanese). Igaku to Seibutsugaku (Medicine and Biology) 93:359–363
Walser M, Bodenlos LJ (1959) Urea metabolism in man. J Clin Invest 38:1617–1626
Levey AS, Madaio MP, Perrone RD (1991) Laboratory assessment of renal disease: Clearance, urinalysis, and renal biopsy. In: Brenner BM, Rector FC (eds) The kidney, 4th edn. W.B. Saunders, Philadelphia, pp 925–929
Mitch WE, Walser M (1991) Nutritional therapy of the uremic patient. In: Brenner BM, Rector FC (eds) The kidney, 4th edn. W.B. Saunders, Philadelphia, pp 2195–2196
Wassner SJ, Li JB (1982) NT-methylhistidine release: Contributions of rat skeletal muscle, GI tract, and skin. Am J Physiol 243:E293–297
Kim YB (1981) Immunology-Introductory remarks. In: Sasaki S, Ozawa A, Hashimoto K (eds) Recent advances in germ-free research. Tokai University Press, Tokyo, pp 483–491
Ueda K, Yamazaki S, Someya S (1973) Impairment and restoration of the delayed type hypersensitivity in germ-free mice. Jpn J Microbiol 17:533–536
Ohwaki M, Yasutake N, Yasui H, Ogura R (1977) A comparative study on the humoral immune responses in germ-free and conventional mice. Immunology 32:43–48
MacDonald TT, Carter PB (1979) Requirement for a bacterial flora before mice generate cells capable of mediating the delayed hypersensivitity reaction to sheep red blood cells. J Immunol 122:2624–2629
Nomoto K (1985) Relation between normal flora and infection, host-defence (in Japanese). Biseibutsu (Microorganism) 1:21–24
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Okada, M. The influence of intestinal flora on wound healing in mice. Surg Today 24, 347–355 (1994). https://doi.org/10.1007/BF02348566
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02348566