The main goal of this study was to obtain normative data of the scores of the Health-Sciences Evidence Based Practice (HS-EBP) questionnaire, and to analyse evidence-based practice (EBP) among potential clusters of osteopathy professionals in Spain. An online descriptive cross-sectional study has been applied. A total number of 443 Spanish practicing osteopaths answered a survey including the 5 dimensions of the HS-EBP questionnaire and sociodemographic, training, and practice variables using the “LimeSurvey” online platform. Results point out that the median scores for each five HS-EBP questionnaire dimensions were 95.00, 86.00, 78.00, 84.00 and 62.00 considering that the range of possible scores in each of the dimensions was: from 12 to 120 in dimensions 1, 4 and 5; from 14–140 in dimension 2; and from 10–100 in dimension 3. A clustering algorithm extracted 6 different profiles across the five HS-EBP latent dimensions: low scores in all dimensions (cluster 1); low scores in all dimensions but with medium scores in dimension 1 (cluster 2); mixed pattern of scores, low in dimensions 2 and 5 and medium in the rest of the dimensions; medium scores in all dimensions (cluster 4); high scores in all dimensions and low scores in dimension 5 (cluster 5); and high scores in all dimensions (cluster 6). Significant relationship was found among the response patterns in the clusters and: academic degree level, EBP training and training level, and work time invested in healthcare activity, research and teaching activity. These results allow a description of the actual level of EBP and differential profiles of Osteopathy care practice in Spain. Knowledge of normative scores of the HS-EBP questionnaire and identification of different predictors of Spanish osteopaths’ EBP, e.g., academic degree, EBP training and training level, work time invested in healthcare activity, research, and teaching activity, and having a working relationship with an accredited educational centre, enable a comprehensive evaluation of the EBP of osteopathic professionals and can also be useful for developing and implementing formative intervention programs for improving EBP practice in osteopathic practice.