Devita Anggraeni
Departemen Ilmu Bedah Dan Radiologi, Fakultas Kedokteran Hewan, UGM

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Keterkaitan Panhisterektomi dan Suplemen 1,25- Dihidroksivitamin D3 dengan Risiko Urolitiasis pada Tikus (CORRELATION BETWEEN PANHISTERCTOMY AND 1.25-DIHYDROXYVITAMIN D3 SUPPLEMENTATION ON RATS UROLITHIASIS RISK) Hartiningsih .; Devita Anggraeni; Irkham Widiyono; Hastari Wuryastuti
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The objective of this research was to study the correlation of panhisterectomy and supplement 1.25-dihydroxyvitamin D3 on urolithiasis risk in Wistar rats. Twenty female Wistar rats at 8 weeks of age, weredivided into four groups (control fed standard diet, control fed standard diet+1,25-dihydroxyvitamin D3 supplement, panhisterectomy fed standard diet and panhisterectomy fed standard diet +1,25-dihydroxyvitamin D3 supplement). Eleven weeks after treatment, each of rats was placed into individualmetabolic cage for balance study for a week. From day 4 to 11 of the balance study, every morning theremaining food, feces, and urine were collected and recorded for calcium (Ca) analysis. At the end ofbalance study, blood samples were taken from canthus retroorbitalis medialis for estrogen analysis. Theresults showed urinary and fecal Ca excretions were not significantly different compared to the controlgroup. Calcium consumption was significantly higher (P<0.05) in panhisterectomized rats compared withthose in control rats. While, estrogen in panhisterectomized group was not significantly different to thosein control rats. Calcium urinary and Ca consumption in rats consuming 1,25-dihydroxyvitamin D3 supplement were significantly higher (P<0.05) compared with those in without 1,25-dihydroxyvitamin D3 supplementation, but Ca excretion in feses was not significantly different. Estrogen in rats consuming1.25-dihydroxyvitamin D3 supplement was significantly lower (P<0.05) compared with the rats that without1,25-dihydroxyvitamin D3 supplemention. It can be concluded that panhisterectomy does not seem to affecturolithiasis risk, while 1,25-dihydroxyvitamin D3 supplement may affect urolithiasis risk. There is likelyno association between panhisterectomy and 1.25-dihydroxyvitamin D3 supplementation on urolithiasisrisk in Wistar rats.
Identifikasi Leptin pada Kesembuhan Luka Tikus yang Diberi Pakan Lemak Tinggi dan Aplikasi Zinc Topikal (LEPTIN IDENTIFICATION ON WOUND HEALING OF RAT GIVEN HIGH FAT DIET AND TOPICAL ZINC APPLICATION) Devita Anggraeni; Dhirgo Adji; Bambang Sutrisno
Jurnal Veteriner Vol 13 No 4 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Leptin is a hormone produced by adipocytes and play an important role in wound healing. The objectiveof this research was to study the role of leptin in wound healing in mice experimentally given high fat dietsand its correlation to zinc. Thirty two male Sprague Dawley rats at three months of age were used in thisstudy. Rats were randomly allotted into four groups (A,B,C and D) of 8. Rats in group A and B were fednormal diet, while rats in group C and D were fed high fat diet. After two months of treatment, skin incisionsurgery was performed at the back side of the rat. Incision wound was closed with single interruptedsuture. The wound of rats in group A and C were treated with vaseline, while in the group B and D weretreated with zinc 10%. One day after surgery, blood sample were collected frpm four rats from each groupand analysed for leptin (Rat leptin ELISA Int, Genway Biotech Inc). Wounded skin from all animals werealso taken for histopathological examination (Haematoxylin and Eosin). Three days after the surgery, thesame analysis were done for the remaining rats. Leptin level was analyzed statistically using ANOVA forfactorial experiment, while histopathologic analysis was done descriptively. The results showed that theleptin level was significantly affected by time (P<0.05), as leptin level in blood at three days after surgerywas significantly lower than the level at one day after surgery. Meanwhile, histopathological examinationshowed that the percentage of epidermal closure in animals in group A,B,C,and D were 75%, 100%, 25%and 75%, respectivelly. Therefore, it was concluded that topical application of zinc might have significanteffect on the wound healing of the rats fed normal diets as well as these that given a high fat diet.
Kombinasi Calcitriol dan Ethynil Ethyl Estradiol Meningkatkan Ekskresi Kalsium Urin dan Risiko Urolitiasis pada Tikus Ovariektomi Hartiningsih Hartiningsih; Ayu Dewi Puspitasari; Nurdysa Diliana Putri; Nurul Arifah; Wari Pawestri; Devita Anggraeni
Jurnal Veteriner Vol 18 No 2 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (173.684 KB) | DOI: 10.19087/jveteriner.2017.18.2.239

Abstract

The high excretion of calcium (Ca) in the urine can trigger the formation of urolith. Estrogen and calcitriol decrease urinary Ca excretion. This study aims to examine the combination of calcitriol and ethinyl ethyl estradiol against Ca urinary excretion and urolithiasis risk of ovariectomized rats. Twentyfive female Wistar rats eight weeks old were divided into five groups: i) normal control (NK); ii) ovariectomized control (OVK); iii) ovariectomized + calcitriol (OVD); iv) ovariectomized + ethinyl ethyl estradiol (OVE); and v) ovariectomized + combination calcitriol and ethinyl ethyl estradiol (OVDE). Seven weeks post-ovariectomy, each rat was put in an individual metabolic cage for the study of Ca balance. At day 4 to 7 of the study, residual feed, urine, and feces were collected daily for Ca analysis. At day 8, the rats were euthanized, the left kidney were collected for histopathological examination. The results showed that combination of calcitriol and ethinyl ethyl estradiol in OVDE rats caused Ca intake and Ca intestinal absorption significantly higher, and urinary Ca excretion tended to be higher although not significantly different compared to OVK rats. Calcium excretion in OVK rat urine was higher compared to the NK rats. The kidney histopathological changes of OVK rats were not different from the NK rats. Histopathological examination of the OVDE group kidney showed protein deposition in the capsular of Bowman’s capsule and proximal tubules, atrophy of the proximal tubules, and necrosis, respectively. It is concluded that the combination of calcitriol with ethinyl ethyl estradiol in ovariectomized rats increased urinary Ca excretion and increased the risk of urolithiasis. ABSTRAK Tingginya ekskresi kalsium (Ca) dalam urin dapat menjadi pemicu terbentuknya urolit. Estrogen dan calcitriol menurunkan ekskresi Ca urin. Penelitian ini dilakukan bertujuan untuk mengkaji kombinasi calcitriol dan ethynil ethyl estradiol terhadap ekskresi Ca dalam urin dan risiko urolitiasis tikus ovariektomi. Sebanyak 25 tikus Wistar betina umur delapan minggu, dibagi menjadi lima kelompok (kontrol normal NK, kontrol ovariektomi OVK, ovariektomi + calcitriol OVD, ovariektomi + ethynil ethyl estradiol OVE, dan ovariektomi + kombinasi calcitriol dan ethynil ethyl estradiol OVDE). Tujuh minggu pascaovariektomi, setiap tikus dimasukan kandang metabolik individu untuk studi keseimbangan Ca. Pada hari ke-4 sampai 7 studi keseimbangan Ca, setiap hari sisa pakan, urin, dan feses dikumpulkan untuk analisis Ca. Pada hari ke-8 studi keseimbangan Ca, tikus dieutanasi, ginjal kiri diambil untuk pemeriksaan histopatologi. Hasil analisis menunjukkan bahwa kombinasi calcitriol dan ethynil ethyl estradiol pada tikus OVDE menyebabkan konsumsi Ca dan absorpsi Ca intestinal nyata lebih tinggi, dan ekskresi Ca dalam urin cenderung lebih tinggi meskipun tidak berbeda nyata dibanding tikus OVK. Ekskresi Ca dalam urin tikus OVK lebih tinggi dibanding tikus NK. Gambaran histopatologi ginjal OVK terlihat tidak berbeda dengan tikus NK. Histopatologi ginjal OVDE terlihat endapan protein dalam ruang kapsuler kapsula Bowman’s dan tubulus proksimalis, tubulus proksimalis atropi, dan nekrosis. Dari hasil penelitian disimpulkan bahwa kombinasi calcitriol dengan ethynil ethyl estradiol pada tikus ovariektomi meningkatkan ekskresi Ca urin dan meningkatkan risiko urolitiasis.
Ekspresi Tartrate-Resistant Acid Phosphatase-5b pada Epifisis Tulang Femur Tikus Ovariektomi yang Mengkonsumsi Calcitriol dan Raloxifene (TARTRATE-RESISTANT ACID PHOSPHATASE-5b EXPRESSION OF EPIPHYSYS DISTALIS FEMUR OVARIECTOMIZED RATS CONSUMING CALCITRIO Hartiningsih .; Devita Anggraeni
Jurnal Veteriner Vol 17 No 1 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Tartrate resistant alkaline phosphatase 5b (TRACP5b) is secreted by osteoclasts during bonedifferentiation and resorption. The objective of the research was to study TRAP5b expression inovariectomized Wistar rat consuming the combinations of calcitriol and raloxifene supplementation foreight weeks. Twenty five female Wistar rats aged eight weeks were randomly divided into five groups:normal control (NK), ovariectomy control (OVK), ovariectomy+calcitriol supplementation (OVD),ovariectomy+ raloxifene supplementation (OVR), and ovariectomy+calcitriol+ raloxifene supplementation(OVDR). At the end of the treatment, blood samples were taken from plexus orbitalis medialis forestrogen analysis. All rats were euthanized, the uteri were taken and weighed. Left femur was taken forhistopatological examination and immunohistochemistry TRAP5b using monoclonal antibody anti TRAP5bwhich was detected with streptavidin-biotin. The results showed that estrogen levels of the rats in OVKgroup were significantly decreased compared to the rats in NK group, meanwhile estrogen levels in the OVDR rat group were significantly decreased compared to the NK and OVK rat group. Histopathologicalobservation of distal femur epiphysis in group NK showed normal structure, meanwhile, distal femurepiphysis in OVK group was found osteoporosis, with some abnormalities, such as: increased of bonemarrow space, domination of adipocytes in the bone marrow, and decrease of trabecular bone speculum inepiphysis. Histopathological findings of distal femur epiphysis in OVDR group were increased of trabecularbone speculum in epiphysis and the domination of adipocytes in the bone marrow of epiphysis.Immunohistochemistry of distal femur epiphysis in OVDR group revealed increasing tartrate resistantalkaline phosphatase 5b (TRAP5b) expression in trabecular bone, which was located in bone marrow spaceand trabecular speculum surface as well. It can be concluded that the combination of calcitriol and raloxifenesupplementation in ovariectomized rats aged 8 weeks increases TRACP5b expression as a marker for theincrease of osteoclast acitivity in bone resorption, which may lead to the formation of the bone.
Respons Tulang Femur Tikus Ovariohisterektomi yang Mengkonsumsi Kasein dan Disuplementasi Calcitriol Selama 30 Minggu (THE RESPONSE OF BONE FEMUR OVARIOHISTERECTOMIZED RATS CONSUMING CASEIN AND CALCITRIOL SUPPLEMENTATION FOR 30 WEEKS) Hartiningsih Hartiningsih; Devita Anggraeni; Irkham Widiyono; Hastari Wuryastuty
Jurnal Veteriner Vol 16 No 1 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Calcitriol supplementation in ovariohisterectomized rat was known to decreased calcium retention.The objective of the research was to study the response of femur bone to calcitriol supplementation for 30weeks in ovariohisterectomized rats consuming casein. Twenty female Wistar rats at 8 weeks of age wererandomly divided into four groups (unovariohisterectomy without calcitriol supplementation (N),unovariohisterectomy with calcitriol supplementation (ND), ovariohisterectomy without calcitriolsupplementation (O) and ovariohisterectomy with calcitriol supplementation (OD) of five each. Thirtyweeks after surgery, femur was taken for histopathological and immunohistochemistry examination.Immunohistochemistry of distal femur metaphysis in group O and OD were revealed decreasing tartrateresistant alkaline phosphatase 5b (TRAP5b) in trabecular bone, which was located in bone marrow space,and also in trabecular speculum surface. Histopathological analysis of distal femur metaphysis in groupN and ND were showed normal structure, meanwhile, distal femur metaphysis in group O and OD wereshown some abonormalities, such as increased of bone marrow space, domination of adipocytes in the bonemarrow, and decreased of trabecular bone speculum in metaphysis. Based on the results, it was concludedthat femur bone of ovariohisterectomized rats fed casein with and without calcitriol 8ng/daysupplementation for 30 weeks were showed unbalanced between resorption and formation of bone whichwas domination by bone resorption.
THE EFFECTIVENESS OF THE COMBINATIONS OF CALCITRIOL AND ETHYNIL ETHYL ESTRADIOL TO DECREASE OSTEOPOROSIS AND ENDOMETRIAL CANCER RISKS IN OVARIECTOMIZED RATS Hartiningsih Hartiningsih; Devita Anggraeni
Jurnal Kedokteran Hewan Vol 11, No 2 (2017): June
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (684.352 KB) | DOI: 10.21157/j.ked.hewan.v11i2.4895

Abstract

The objective of this research was to study the effectiveness of calcitriol and ethynil ethyl estradiol combinations to decrease osteoporosis and endometrial cancer risks in ovariectomized Wistar rats. Twenty five 8-week old female Wistar rats were randomly divided into five groups (normal controlled rats/NK, ovariectomized rats/OVK, ovariectomized rats + calcitriol supplementation/OVD, ovariectomized rats + ethynil ethyl estradiol supplementation/OVE, and ovariectomized rats + calcitriol + ethynil ethyl estradiol supplementation/OVDE). At the end of the study, all rats were weighed and euthanized with 10% ketamine and 2% xylazine. Uterus and left femur were taken and fixed in 10% formaldehyde solution for histopathological examination using hematoxylin and eosin stain. Uterus was weighed before the fixation. The results showed that the percentage of uterus weight in OVK was significantly decreased compared to NK. Meanwhile, the percentage of uterus weight in OVDE was significantly increased compared to OVK. Histopathological features of the uterus in OVK were atrophy, reduction of myometrial and endometrial layers thickness, and formation of cuboidal epithelium in the endometrial lumen. However, the thickness of myometrial and endometrial layers in OVDE were increased, and its endometrial lumen was lined with metaplastic and hyperplastic squamous cell. Histopathological features of distal femur epiphysis in OVK ras showed fewer trabecular spiculums and more adipocyte in the bone marrow compared to NK. However, OVDE had more trabecular bone spiculum and less adipocyte in the bone marrow compared to OVK. In conclusion, the combination of calcitriol and ethynil ethyl estradiol supplementation could reduce osteoporosis, but increased the endometrial cancer risk in ovariectomized Wistar rats.
EFEKTIVITAS DIET TERI TAWAR DAN SUPLEMENTASI KALSITRIOL UNTUK MENCEGAH OSTEOPOROSIS PADA TIKUS OVARIEKTOMI The Effectiveness of Fresh Anchovies Diet and Calcitriol Supplementation to Prevent Osteoporosis in Ovariectomized Rats (The Effectiveness of Fresh Anchovies Diet and Calcitriol Supplementation to Prevent Osteoporosis in Ovariectomized Rats) Hartiningsih H; Devita Anggraeni; Sudarminto S
Jurnal Kedokteran Hewan Vol 10, No 1 (2016): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (458.007 KB) | DOI: 10.21157/j.ked.hewan.v10i1.3362

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This study was conducted to assess the effectiveness of a diet contained fresh anchovies and calcitriol supplementation for 6 weeks to prevent osteoporosis in ovariectomized rats. Fifteen Wistar rats aged 8 weeks were divided randomly into 3 groups (normal/K, ovariectomized/Ov, and ovariectomized + calcitriol/OVD), 5 mice each. Group K and Ov rats were fed with fresh anchovies, while the OVD group was fed with fresh anchovies + calcitriol. At the age of 15 weeks, all mice were done for euthanasia, then left femur was collected for immunohistochemistry examination of tartrate resistant acid phosphatase5b (TRAP5b). The detection of (TRAP5b) was conducted using a monoclonal ant ibody anti TRAP5b, and detected using a streptavidin-biotin. The results showed that the metaphysis part of distal femur bone of mice group K, Ov, and OVD were positive TRAP5b stained with brown color on trabecular bone in bone marrow cavity and trabecular spiculum surface, but rats in group K and Ov had extensive bone marrow cavity and normal trabecular spiculum, whereas OVD group showed bone marrow cavity dilation, accumulation of adipocytes in the bone marrow cavity, and shorter the spiculum of trabeculae. It can be concluded that fresh anchovies diet and calcitriol supplementation for 6 weeks are not effective in preventing osteoporosis in ovariectomized rats.Key words: calcitriol, femur, ovariohisterectomy
KESEMBUHAN LUKA SETELAH PEMBERIAN TOPIKAL ZINK PADA TIKUS DENGAN PAKAN LEMAK TINGGI Devita Anggraeni; Dhirgo Adji; Retno Murwanti
Jurnal Kedokteran Hewan Vol 9, No 2 (2015): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (454.192 KB) | DOI: 10.21157/j.ked.hewan.v9i2.2809

Abstract

Penelitian ini bertujuan mengetahui peran zink pada kesembuhan luka dalam kondisi berat badan berlebih. Dalam penelitian digunakan 20 ekor tikus Sprague Dawley jantan berumur tiga bulan (150-200 g). Tikus dibagi secara acak menjadi empat kelompok (A, B, C, dan D), masing-masing kelompok terdiri atas lima ekor. Kelompok A dan B diberi pakan normal, sedangkan kelompok C dan D diberi pakan lemak tinggi. Setelah dua bulan perlakuan pakan, seluruh tikus dioperasi untuk membuat luka irisan pada kulit, kemudian luka ditutup dengan jahitan. Luka jahitan pada kelompok A dan C diberi aplikasi topikal unguenta vaseline album (tanpa zink), sedangkan luka jahitan pada kelompok B dan D diberi aplikasi topikal unguenta zink 10% dengan bahan dasar vaselin album. Lima hari setelah operasi, seluruh tikus diambil darahnya untuk dianalisis terhadap jumlah leukosit total kemudian diikuti pengambilan jaringan kulit untuk pemeriksaan histopatologis dan penghitungan jumlah leukosit di jaringan. Hasil penelitian menunjukkan adanya interaksi yang signifikan antara pakan dan aplikasi topikal terhadap jumlah leukosittotal dalam darah (P0,05). Jumlah leukosit di jaringan mengalami penurunan setelah aplikasi topikal zink, meskipun hasilnya tidak signifikan (P0,05). Analisis histopatologis menunjukkan epidermis sudah tertutup pada seluruh tikus perlakuan, namun demikian sebagian tikus masih menunjukkan dermis yang terbuka, yaitu kelompok A sebanyak 60%, kelompok B sebanyak 60%, kelompok C sebanyak 60%, dan kelompok D sebanyak 40%. Dapat disimpulkan bahwa proses kesembuhan luka setelah pemberian topikal zink pada tikus yang diberi pakan lemak tinggi menunjukkan hasil yang baik, yaitu ditandai dengan penutupan lapisan epidermis dan dermis, serta menurunnya jumlah leukosit di jaringan.
THE EFFECTIVENESS OF ETHANOL EXTRACT OF BINAHONG LEAVES ON DIABETIC WOUND HEALING Devita Anggraeni; Claude Mona Airin; Slamet Raharjo
Jurnal Kedokteran Hewan Vol 11, No 4 (2017): December
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (639.445 KB) | DOI: 10.21157/j.ked.hewan.v11i4.6562

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This research aimed to study the effectiveness of ethanol extract of binahong leaves on blood glucose, insulin, blood chemical profiles (serum glutamic pyruvate transaminase=SGPT, serum glutamic oxaloacetic transaminase=SGOT, ureum, and creatinine), and skin histopathology in diabetic rat. A total of 20 male Wistar rats aged 3 months (± 250 gram) were divided into five groups, with four rats in each group. Group I (non-diabetic control) was injected with 0.1 M sodium citrate buffer, while group II (diabetic control), III, IV, and V were injected with single dose of Streptozotocin (STZ) at dose 40 mg/kg intraperitoneally (IP). One week after the injection, the dorsal skin of the rats were excised. Group I and II were given cream topically and 1% NaCMC orally, group III was given 50% ethanol extract of binahong leaves (EEB) topically and 1% NaCMC orally, group IV was given cream topically and EEB 300 mg/kg orally, and group V was given 50% EEB topically and EEB 300 mg/kg orally. These treatments were continued for 14 days. Blood samples were obtained at the end of study to examine blood glucose, insulin, and blood chemical profiles (SGOT, SGPT, ureum, and creatinine). Examination of skin histopathology and leukocyte count were also done. The result showed that blood glucose, insulin, SGOT, SGPT, and ureum level of diabetic rats given topical or oral EEB did not significantly different from diabetic control group, even though blood glucose, insulin, SGOT, SGPT, and ureum level of diabetic rats given topical and oral EEB were found lower compared to diabetic control group. Administration of EEB 300 mg/kg orally in diabetic rats could lower creatinine level significantly (P0.05). Histopathological examination of dorsal skin of diabetic rats which were given EEB topically showed the decrease of fibroblast proliferation, leukocyte infiltration, and hemorrhage in dermis area. Leukocyte count on skin tissue was significantly lower (P0.05) in diabetic rats given EEB. In conclusion, topical or oral administration of EEB can help healing process in diabetic wound.
THE EFFECTIVENESS OF THE COMBINATIONS OF CALCITRIOL AND ETHYNIL ETHYL ESTRADIOL TO DECREASE OSTEOPOROSIS AND ENDOMETRIAL CANCER RISKS IN OVARIECTOMIZED RATS Hartiningsih Hartiningsih; Devita Anggraeni
Jurnal Kedokteran Hewan Vol 11, No 2 (2017): June
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i2.4895

Abstract

The objective of this research was to study the effectiveness of calcitriol and ethynil ethyl estradiol combinations to decrease osteoporosis and endometrial cancer risks in ovariectomized Wistar rats. Twenty five 8-week old female Wistar rats were randomly divided into five groups (normal controlled rats/NK, ovariectomized rats/OVK, ovariectomized rats + calcitriol supplementation/OVD, ovariectomized rats + ethynil ethyl estradiol supplementation/OVE, and ovariectomized rats + calcitriol + ethynil ethyl estradiol supplementation/OVDE). At the end of the study, all rats were weighed and euthanized with 10% ketamine and 2% xylazine. Uterus and left femur were taken and fixed in 10% formaldehyde solution for histopathological examination using hematoxylin and eosin stain. Uterus was weighed before the fixation. The results showed that the percentage of uterus weight in OVK was significantly decreased compared to NK. Meanwhile, the percentage of uterus weight in OVDE was significantly increased compared to OVK. Histopathological features of the uterus in OVK were atrophy, reduction of myometrial and endometrial layers thickness, and formation of cuboidal epithelium in the endometrial lumen. However, the thickness of myometrial and endometrial layers in OVDE were increased, and its endometrial lumen was lined with metaplastic and hyperplastic squamous cell. Histopathological features of distal femur epiphysis in OVK ras showed fewer trabecular spiculums and more adipocyte in the bone marrow compared to NK. However, OVDE had more trabecular bone spiculum and less adipocyte in the bone marrow compared to OVK. In conclusion, the combination of calcitriol and ethynil ethyl estradiol supplementation could reduce osteoporosis, but increased the endometrial cancer risk in ovariectomized Wistar rats.