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A novel hydroxyapatite (HAp)/poly(L-lactic acid) (PLLA) nanocomposite nonwoven fabric, which was coated and mixed with calcined HAp nanocrystals, and has submicron-sized dimples on its surface, was fabricated. First, HAp-mixed PLLA fabric was prepared by electrospinning a HAp nanocrystal dispersion in dichloromethane (DCM)-dissolved PLLA. It was found that most of the HAp nanocrystals were not exposed on the HAp-mixed PLLA fiber surface but covered with the PLLA matrix. A HAp-nanocrystal coating was applied onto the surface of the HAp-mixed PLLA fabric after corona discharge treatment followed by ethanol washing. The submicron-sized dimples were enlarged after the ethanol washing. After the HAp-nanocrystal coating, the HAp-mixed PLLA fabric surface was uniformly coated with the HAp nanocrystals. In vitro cell spread tests showed that the rat osteoblasts spread more on HAp-nanocrystal-coated fabrics than on non-HAp-coated fabrics. Upon covering calvarial defects, the in vivo hard tissue responses suggested earlier restoration of the defects with HAp-nanocrystal-coated fabrics than those with non-HAp-coated fabrics.

ABSTRACT. Serum alpha-fetoprotein (AFP) values were measured in hepatic diseased dogs with or without tumor and non-hepatic tumor bearing dogs by a sandwich ELISA using anti-dog AFP antiserum. Serum AFP values were less than 70 ng/ml in clinically healthy dogs. The values in dogs with hepatocellular carcinoma were higher than 1,400 ng/ml in 7 of 9 dogs, wherever those in two dogs with cholangiocarcinoma were in the normal range. Serum AFP values in hepatic diseased dogs without tumor were also high, however, the values were below 500 ng/ml in 90% of the dogs. In non-hepatic tumor dogs, serum AFP values were less than 500 ng/ml in 76% of the dogs. In the surgically removal cases with hepatocellular carcinoma, serum AFP values rapidly decreased. These results suggested that the sandwich ELISA using anti-dog AFP antiserum was an available method for diagnosis of hepatocellular carcinoma in dogs.-KEY WORDS: alpha-fetoprotein, canine, ELISA, tumor.J. Vet. Med. Sci. 61(6): 657-659, 1999 with hepatocellular carcinoma were also monitored AFP values after surgical removal of masses. Serum: Sera were collected by the usual methods, and kept below 20°C until assay.Preparation of AFP and antibodies: Canine AFP and antidog AFP antiserum were prepared according to the previously described method [17].ELISA for canine AFP: The protein concentration of purified AFP was measured by the Coomassie protein assay methods (Pierce, U.S.A.) using bovine serum albumin (BSA) as a standard. The ELISA for canine AFP was developed by the previously reported avidin-biotin sandwich method with minor modifications [17]. Briefly, the immunoglobulin G (IgG) fraction of anti-dog AFP antiserum from rabbits was coated on 96-well microplates (flat-bottom) at 5 µg per well. The wells were blocked with PBS containing 0.5% BSA, and washed with PBS containing 0.05% Tween 20 (TPBS) by an ELISA plate washer (Immunowash; BioRad, USA). Serum samples were diluted with TPBS at 1:2, 1:10, and 1:50. A standard solution of canine AFP was also diluted at concentrations of 0.25 ng/ml to 600 ng/ml in TPBS. The IgG fraction of anti-dog AFP antiserum was labeled with N-hydroxysuccimidobiotins (Sigma, U.S.A.) as shown by Coligan et al. [3]. The peroxidase-conjugated streptavidin was purchased from Dako (Denmark) and the chromogen 2',2' azino-bis (3-ethylbenzthiazolin-6-sulfonic acid) from Sigma (U.S.A.). Absorption of the samples and standards were measured at 405 nm. Serum AFP was measured in less than 3 weeks after storage, because storage time would affect AFP values [7]. RESULTS Serum AFP values:Serum AFP values in clinically healthy dogs were less than 70 ng/ml. Serum AFP values Serum alpha-fetoprotein (AFP) values have been used for the diagnosis of hepatocellular carcinoma [4,[5][6]14] and also for the understanding pathophysiological states in hepatic failure [8][9] in man. The values in dogs measured by enzyme-linked immunosorbent assay (ELISA) or radio immuno-assay (RIA) using anti-human AFP antibody were also reported [7,9,15], as the incidence of hepato...

Abstract. Serum alpha-fetoprotein (AFP) concentrations were measured before and after surgical removal of tumor masses in four dogs with hepatocellular carcinoma (HCC). Localization of AFP was also examined immunohistochemically in tumor tissues. In three cases, the serum AFP concentration was 10 to 20 times higher than that of normal dogs. One to two months after surgery, the serum AFP concentration had decreased to normal range. AFP was localized in the tumor tissues in these three cases. One case, which had a low serum AFP, did not show AFP localization in tumor tissue.