Effect of Vitamins D3 and K2 in uOc and Insulin Serum Levels in Patients With Diabetes Mellitus
Condition: Diabetes Mellitus, Type 2; Bone Loss; Insulin Resistance
MeSH terms: Diabetes Mellitus; Diabetes Mellitus, Type 2; Insulin Resistance
Study type: Interventional

Summary: 
      Introduction: Patients with DM2 have chronic hyperglycemia derived from a decrease in insulin
      sensitivity, cause of comorbidities such as bone demineralization, decreasing quality of life
      and increasing mortality. This could be related to changes in the serum levels of
      carboxylated Osteocalcin and Insulin, together with the deficit the daily consumption of
      vitamins D3 and K, which is crucial for the process of mineralization of the bone matrix.

      Research question: What is the effect of supplementation with Vitamins D3 and K2 on serum
      levels of Carboxylated Osteocalcin and Insulin in patients with Type 2 Diabetes mellitus?

      Hypothesis: Supplementation with Vitamins D3 and K2 modifies the serum levels of Carboxylated
      Osteocalcin and Insulin in patients with Type 2 Diabetes mellitus.

      General Objectives: To assess the effect of supplementation with Vitamins D3 and K2 on serum
      levels of Carboxylated and Non-Carboxylated Osteocalcin in patients with Type 2 Diabetes
      mellitus.

      Material and Methods: Clinical trial, double blind, randomization, 40 patients with DM2,
      35-65 years, supplementation (3 months), clinical and laboratory determinations (uOC and
      Insulin).

        -  Group 1: Vitamin D3 1000UI + Placebo

        -  Group 2: Vitamin K2 100 mcg + Placebo

        -  Group 3 (Positive Control): Vitamins D3 1000UI + K2 100 mcg
    

Detailed description: 
      Type 2 diabetes mellitus is a group of metabolic disorders characterized by chronic
      hyperglycemia, being one of the main causes of mortality in our country.

      There are different comorbidities, including bone demineralization, which involve an
      imbalance in the process of bone formation and resorption, which entails the risk of
      decreasing bone mineral density. These processes could be related to the serum levels of
      Non-carboxylated Osteocalcin and Insulin, said molecule locally modulates the process of bone
      mineralization.

      On the other hand, it is known that the daily consumption of vitamins in a general diet in
      our population (specifically including vitamins D3 and K), is below the necessary basic
      requirements, so considering crucial elements for the benefit of Bone matrix mineralization,
      which is why the present study proposes supplementation with the aforementioned vitamins in
      patients with Type 2 DM, to avoid a decrease in bone mineral density, increasing the
      mortality rate of individuals suffering from it.

        1. - The study subjects from the West of the country were invited to participate in the
           research protocol, where their objective was explained and the signing of the consent
           was requested under information, where a structured evaluation was subsequently carried
           out consisting of :

           a) Comprehensive clinical evaluation (including general sociodemographic data of a
           complete medical history).

        2. - Once the previous data was obtained, the randomization of the treatment was carried
           out, classifying the patients in the following groups:

             -  Group 1 Vitamin D3 1000UI + Placebo (Calcined Magnesia).

             -  Group 2 Vitamin K2 100 mcg + Placebo (Calcined Magnesia).

             -  Group 3 (Positive Control) Vitamins D3 1000UI + K2 100 mcg.

        3. - Blood sample collection through Punzocat, which was placed in a red tube, for
           subsequent centrifugation at 3500rpm x 15 minutes, performing aliquots of the serum
           obtained and stored at -80 ° C and thus perform the pertinent quantifications of the
           molecules to study.

        4. - Quantification of serum levels of non-carboxylated Osteocalcin and Insulin

           a) The determinations corresponding to the serum levels of Non-carboxylated Osteocalcin
           and Insulin were made, both baseline prior to the intervention as well as the final
           quantification, once the supplementation was finished, by means of the indirect ELISA
           technique (commercial reagents -Takara were used - for the realization of these
           determinations), which involved the following process: In a plate the wells were coated
           with a first antibody, then a wash was performed to remove the excess antibody, after
           that, the sample in which the antigen was found was added, which was retained in the
           well after being recognized by the first antibody, a second wash was subsequently made
           to remove unbound material, then a solution with a second labeled anti-antigen antibody
           was applied. Thus each antigen molecule was bound to an antibody in the base that kept
           it fixed and a second antibody for its reaction and labeling. Finally, the plate was
           introduced into a spectophotometer and the absorbance was measured at a length of 450
           nm, for both non-carboxylated Osteocalcin and Insulin.

        5. - The quantification in serum of cholesterol and triglycerides was carried out by using
           the fully automated random access analyzer equipment for clinical chemistry, model ERBA
           XL 200.

        6. - The evaluation of the HOMA index was calculated by using the equations HOMA-IR and
           HOMA-B, for insulin resistance (IR) and Percentage, respectively.

        7. - The daily of adherence to treatment was provided, so that they recorded all those
           important events that would ensure adequate intake of vitamins, as well as adverse
           reactions to them, which were evaluated in scheduled appointments, in addition to each
           The corresponding bottles were delivered consecutively for each of the 3 months of
           intervention, with the corresponding vitamins for each patient according to their
           allocation group.

        8. - Finally, in the last visit the results obtained during the whole study were made known
           to each of the patients.
    

Intervention name(s): Supplementation with Vitamin D3 y K2
Intervention type(s): Dietary Supplement
Intervention description(s): Supplementation at a dose of 1 capsule every 24 hours x 3 months
Intervention MeSH terms: Vitamin D; Cholecalciferol
Allocation: Randomized
Intervention model: Single Group Assignment
Intervention model description: Controlled clinical trial, with randomization simple, double blind.
Masking: Triple (Participant, Care Provider, Investigator)
Primary purpose: Supportive Care
Keywords: diabetes; osteocalcin; insulin